0001] The present application claims priority to United States Provisional

Patent Application No. 63/352,468 filed on June 20, 2016, the entirety of which is incorporated herein by reference.

FIELD OF THE INVENTION

[0002] The present disclosure relates to the field of cell processing employing automated systems and, more particularly, relates to apparatus and method for processing cells for use in cell therapy and regenerative medicine, as well as other biological samples.

BACKGROUND

[0003] Stem cell therapies hold much promise for regenerative medicine.

Stem cells have the potential to develop into many different cell types in the body and can theoretically divide without limit to replenish cells in need of repair. There are different types of stem cells with varying ranges of commitment options. Embryonic stem cells hold great potential for regenerative medicine, however, they have many disadvantages including the possibility of transplant rejection and possible teratoma formation if the cells are not properly differentiated prior to transplantation. Adult stem cells such as neural stem cells (NSC) and oligodendrocyte precursor cells (OPC) have a more restricted developmental potential than embryonic stem cells and generally differentiate along their lineage of origin. While adult neural stem cells also represent a promising treatment option for neurodegenerative disorders, there are numerous disadvantages, including difficulty of isolation, limited expansion capability, and immune rejection of transplanted donor cells. The same or similar limitations apply for most other cells and stem cells.

[0004] For a stem cell to graft permanently and efficiently (in a functional manner) into a patient's tissue, the stem cell is ideally autologous (i.e., the patient's own). There is a desire therefore in the medical, scientific, and diagnostic fields to reprogram an easily obtainable cell (such as a somatic cell) from a patient into a stem- like cell, preferably without fusing or exchanging material with an oocyte or another stem cell, for use in stem cell therapy. Methods for generating safe and efficacious autologous stem cells for a specific tissue, organ or condition to be treated, as well as new stem cells with new or unique features such as enhanced potency and/or safety, have been reported. For example, Ahlfors et al. describe methods of reprogramming easily obtainable cells to highly desirable multipotent or unipotent cells, including stem-like cells and progenitor-like cells as well as cell lines and tissues, by a process of in vitro dedifferentiation and in vitro reprogramming (International PCT Application Publication No. WO2011/050476, U.S. Patent Application Publication Nos. US20120220034, US20120288936, and US20140038291). Such cells can potentially be transplanted back into a patient to regenerate damaged or lost tissue in a wide range of disorders and conditions such as Parkinson's disease, multiple sclerosis, heart disease, spinal cord injury, cancer, and so on.

[0005] However, the use of such cells in human therapy is severely restricted by the limitations of current production methods which are long, labor-intensive, inefficient, and expensive. Realizing the full potential of cell therapies, especially autologous stem cell therapies, will require addressing the challenges inherent in obtaining appropriate cells for millions of individuals while meeting the regulatory requirements of delivering therapy and keeping costs affordable. It is estimated that, using current production methods for iPS cells (induced pluripotent stem cells) or reprogrammed cells, two people working in a single clean room can only process about 20 samples per year, assuming that no samples are lost due to bacterial or cross contamination or human error, and the costs of production are prohibitive. In addition to this, several quality control personnel are needed to determine the identity, purity, potency, etc. of the cells as well as ensuring the cell product is not contaminated. Many of these same challenges and requirements apply for producing or maintaining various cell lines, e.g., for research purposes, as well as for producing biological products or biomaterials where cells or tissues are involved.

[0006] Generally, with current production methods, only one cell-line can be processed at a time to ensure no risk of cross-contamination, and equipment must be sterilized between each sample. It may take weeks or months to process one cell line. In order to meet Good Manufacturing Practices (GMP) guidelines e.g., for human

somatic cell therapy, all steps must be performed in a clean room meeting CLIA or other requirements and in the presence of at least two persons. Multiple complex and precisely-timed steps must be performed, along with safety testing and analytical testing for quality control throughout, all of which must be documented in detail. Cells must also meet stringent safety and potency standards for approval for human therapeutic use. Clearly there is a need for improved methods of generating specific cells suitable for particular human therapeutic applications especially from autologous human cells and other types of cells, in particular to increase the speed and efficiency of cell processing and quality control analysis while reducing the risk of cross-contamination between cell lines and the risk of human error, in order to meet regulatory guidelines and at affordable cost.

[0007] U.S. Patent No. 8,784,735 describes an apparatus for automated processing of biological samples. There is described an apparatus for automated processing of at least one biological sample accommodated on a carrier member, such as a slide, by applying a predetermined amount of reagents in a predetermined sequence according to a processing protocol, said apparatus comprising: a housing frame; at least one processing section for accommodating at least one slide, the at least one processing section being provided within the housing; a hood cover protecting the at least one processing section in said housing, wherein the hood cover completely encloses the processing section defining an interior space; and wherein the apparatus further comprises a climate control device provided to control the environment within the interior space. While the disclosed apparatus and methods are suitable for processing fixed biological samples, they cannot be used to process live biological samples such as dividing cells and cell lines.

[0008] Commercially available cell culture processing systems such as

Cellmate™ (Sartorius Stedim, Wilmington, DE, U.S.A.) provide full automation of processes needed to culture cells in roller bottles and T-flasks. Such systems offer large volume, single cell-line production including automated cell seeding, enzymatic and mechanical harvesting, cell sheet rinsing, media changing, and transient transfection. The Cellmate™ system was developed for a GMP environment. However, such systems can only be used in a clean room and can only process one cell-line at a time, as they do not control for cross-contamination between cell lines.

They are not fully automated, still requiring human handling for certain steps or functions (such as capping and uncapping tubes) and other analytical assays. Although the Cellmate™ system can measure cell count, cell viability, and cell confluency, it cannot perform other quality control tests needed to meet GMP regulations (such as tests for identity, potency, purity, sterility, etc.).

[0009] CompacT SelecT™ (Sartorius Stedim, Wilmington, DE, U.S.A.) provides an automated cell culture system for maintaining and expanding multiple cells lines, including plating cells ready for assaying, harvesting cells, performing transfections, and determining cell number and viability. The system includes a flask incubator, an aseptic processing environment, and various plating modules, along with bar-coded tracking. However, the system can only be used in a clean room and can only process one cell-line at a time, as it does not control for cross-contamination between cell lines. The system is suitable only for expanding cells, not for processing of cells (such as reprogramming) and cannot perform quality control tests needed to meet GMP regulations. The system is not fully automated, still requiring human handling for certain steps or functions. For example, in order to reload supplies into the system, it must be manually opened and re-stocked.

[0010] Fulga et al. (U.S. Patent Application Publication No. 2011/0206643) describes an automated cell processing system for receiving a tissue containing a multiplicity of cells belonging to multiple cell types, and automatically increasing both the proportion and the absolute number of cells of at least one of the multiple cell types as compared with at least another of the multiple cell types. A self- scraping cell culture assembly comprising a generally annular dish defining a generally flat, circularly- shaped cell growth surface; a cover arranged for sealing engagement with the annular dish; and at least one scraper blade mechanically associated with the cover, whereby rotation of the cover relative to the dish provides scraping of cells from the circularly- shaped cell growth surface. The system also includes an automated packaging functionality. However, the system is not fully automated and has many of the limitations of other systems described above.

SUMMARY

[0011] It is an object of the present invention to ameliorate at least some of the inconveniences present in the prior art.

[0012] There are provided herein systems and methods for automated processing of biological samples that are executable without handling by a human operator and/or are capable of processing a plurality of batches at the same time without cross -contamination between batches, optionally under conditions that meet GMP guidelines and regulations.

[0013] In some implementations, systems are designed to maintain sterility to such an extent that they need not be operated in a clean room. For example, the system can be restocked with consumables such as reagents, media, plasticware and the like without disturbing the sterility of the system or exposing the system to the outside environment. In some implementations, systems can perform Quality Control (QC) tests such as verifying cell identity, cell purity, cell potency, and/or batch sterility (i.e., no contamination), during or after processing. In some implementations, end-to-end processing is provided, i.e., a biological sample is introduced into the system and the desired end product is presented by the system after processing, without requiring handling by a human operator. In some implementations, monitoring, tracking and recording systems keep detailed records of every step of the process, including QC testing. Such records can be used for quality assurance purposes and to verify that all applicable regulations have been met. In some implementations, quality assurance (QA) of the end product and/or end product release is performed without requiring a human operator. In some implementations, the product is stored and/or packaged for transport after completion of QC and QA without requiring a human operator.

[0014] In some implementations, therefore, systems and methods described herein may provide one or more of the following advantages: allowing processing of multiple biological samples or batches in sequence or at the same time without cross-contamination between samples/batches and/or under GMP conditions (conditions that meet Good Manufacturing Practices (GMP) guidelines or regulations); allowing fast, efficient, and/or affordable processing; being executable without human intervention during the processing (except to restock consumables, which can be done without interrupting processing or disrupting sterility / the aseptic environment); providing fully automated end-to-end processing, that may also include storage and/or packaging of the final end product; obviating the need for personnel operating in a clean room e.g., meeting CLIA requirements; having integrated analytical and quality control (QC) capabilities, including all QC testing required for GMP guidelines and regulations; providing detailed reports of the processing for quality assurance purposes; and verifying automatically that the end product meets applicable regulations and is suitable for its intended purpose, such as human therapy. In some implementations, systems and methods described herein provide increased efficiency and quality of processing over previous systems.

[0015] Systems and methods may be used for a wide variety of processing on many different types of biological samples. For example, systems and methods may be used to reprogram or transform cells of a first type (such as somatic cells, stem cells, progenitor cells) to cells of a desired second type (such as multipotent, unipotent, or pluripotent cells) for use e.g. in human therapy. Systems and methods may be used for direct reprogramming of cells; for production of multipotent, unipotent, or pluripotent cells; for production of stem-like or progenitor- like cells; for production of induced pluripotent stem cells (iPSCs); for production of embryonic stem cells; and for production of other cells useful for therapeutic, diagnostic, or research purposes. Methods of in vitro dedifferentiation and in vitro reprogramming are detailed in, for example, International PCT Application Publication No. WO2011/050476, U.S. Provisional Application No. 61/256,967, U.S. Patent Application No. 14/958,791, and U.S. Patent Application Publication Nos. US20120220034, US20120288936, and US20140038291, all of which are hereby incorporated by reference in their entirety. Systems and methods may also be used for growth or expansion of cells; for transfection of cells, including stable transfection; for gene editing, including gene insertion, gene deletion, and gene correction; for treatment of cells, e.g., with compounds, antibodies, or other active agents; for inducing differentiation of cells; and combinations thereof. Cells may be manipulated or treated before, during, or after expansion depending on the starting number of cells and the desired end product. Systems and methods may also be used for generation of biomaterials (e.g., tissues, matrices, etc.), generation of biologies (e.g., proteins, antibodies, vaccines, growth factors, etc.), processing of tissues into single cells and/or extraction of extracellular matrix components, for growth of tissues, and for

growth or expansion of cells and cell lines, as well as for screening or discovery research. For example, systems and methods may be used to express and purify therapeutic proteins, antibodies, growth factors, and the like; produce a tissue matrix from a blood sample; isolate and expand a desired cell type from a population of cells; purify extracellular matrix components; expand a cell line; differentiate cells; reprogram or transform cells; transfect cells to introduce vectors, plasmids, RNAs, therapeutic molecules, and the like; repair genetic mutations in cells; and so on. It is contemplated that other applications for processing a product or determining an end product are possible and neither the type of processing nor the type of biological sample being processed is meant to be particularly limited. As used herein, the term "processing" is meant to encompass broadly any such modification, extraction, purification, maintenance, production, expression, growth, culturing, transformation, expansion or treatment of biological samples, particularly live biological samples such as dividing cells and cell lines and tissues containing dividing cells and cell lines. In certain implementations, a "biological sample" does not include samples that have been treated with a fixative agent, e.g., for histological examination.

[0016] In a first broad aspect, there is provided a system for automated processing of batches, the batches being derived from biological samples, the system comprising: a closed and sterile (i.e., aseptic) enclosure; a plurality of reagent containers; at least one reagent dispenser; a quality control module for analyzing at least one characteristic of a batch; a harvesting module; a robotic module; and a control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling the automatic processing of the batches, the automatic processing being executable without handling by a human operator. The system may further comprise numerous components, modules, processing stations, etc., as described herein. In some implementations, the enclosure is at least a Class 10 or ISO 4 environment. In some implementations, the system is configured to automatically process a plurality of batches. In some implementations, the system is configured to automatically process the plurality of batches in compliance with good manufacturing practice (GMP) regulations or guidelines, i.e., under GMP conditions. In some implementations, at least one of the quality control module, the harvesting module, and the robotic module is housed inside the enclosure, automatic processing of cells being conducted inside the enclosure.

[0017] In a second broad aspect, there is provided a system for automated processing of a plurality of batches, the batches being derived from biological samples, the system comprising: a closed and sterile (i.e., aseptic) enclosure; a plurality of reagent containers; at least one reagent dispenser; a quality control module for analyzing at least one characteristic of a batch; a harvesting module; a robotic module; and a control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling the automatic processing of the batches, the system being configured to automatically process the plurality of batches without cross -contamination between batches. In some implementations, the system is configured to automatically process the plurality of batches at the same time using sequential processing. In some implementations, the system is configured to automatically process the plurality of batches in compliance with good manufacturing practice (GMP) regulations or guidelines, i.e., under GMP conditions. In some implementations, the automatic processing is executable without handling by a human operator. The system may further comprise numerous components, modules, processing stations, etc., as described herein. In some implementations, the enclosure is at least a Class 10 or ISO 4 environment. In some implementations, at least one of the quality control module, the harvesting module, and the robotic module is positioned inside the enclosure, automatic processing of cells being conducted inside the enclosure.

[0018] In some implementations, systems described herein further comprise an isolator, the enclosure being selectively fluidly connected to the isolator, and objects from outside the system being received into the enclosure via the isolator, objects from inside the enclosure being passed out of the system via the isolator. In some implementations, the system further comprises a biological safety cabinet (BSC), the isolator being selectively fluidly connected to the BSC, and objects from outside the system being received into the isolator via the BSC, objects from inside the enclosure being passed out of the system by passing from the enclosure to the isolator and from the isolator to the BSC via the isolator.

[0019] In some implementations, two or more systems are selectively fluidly connected to each other, e.g., via an incubator, a freezer, or other similar component disposed outside the enclosures and selectively fluidly connected to each enclosure or system.

[0020] In a third broad aspect, there is provided an automated method for processing a batch in a closed and sterile (i.e., aseptic) enclosure, the batch being derived from a biological sample inserted into the enclosure, the automated method comprising: automatically processing the batch with one or more reagents; automatically analyzing at least one characteristic of the batch; and after automatically processing the batch, automatically harvesting the batch for reception outside the enclosure; the automated method being executable without any handling by a human operator. In some implementations the batch comprises a plurality of batches, and the method comprises automatically processing each of the plurality of batches without cross-contamination between batches. In some implementations, the method is executed in compliance with good manufacturing practice (GMP) regulations and guidelines, i.e., under GMP conditions, and/or in a class 10 environment.

[0021] In a fourth broad aspect, there is provided an automated method for processing a batch in a closed and sterile (i.e., aseptic) enclosure, the batch being derived from a biological sample inserted into the enclosure, the automated method comprising: automatically processing the batch with one or more reagents; automatically analyzing at least one characteristic of the batch; and after automatically processing the batch, automatically harvesting the batch for reception outside the enclosure; wherein the automated method is capable of processing a plurality of batches without cross-contamination between batches. In some implementations, the plurality of batches are processed at the same time using sequential processing. In some implementations, the plurality of batches are processed in compliance with good manufacturing practice (GMP) guidelines, e.g., under GMP conditions. In some implementations, the automated method is executable without any handling by a human operator.

[0022] In some implementations, methods provided herein further comprise quality control (QC) testing during and/or after processing, such as tests for identity, potency, purity, and sterility. In some implementations, methods provided herein further comprise analytical and/or diagnostic testing, such as determination of cell number, viability, and confluency, presence or absence of specific cell markers, growth or differentiation profile, activity, detection of gene mutations, and the like. In some implementations, methods provided herein further comprise monitoring, tracking and/or recording details of every step of the process, including QC testing, for quality assurance purposes and to verify that all applicable regulations have been met.

[0023] In some implementations, systems and methods provided herein include functionalities which expand cells and which conduct quality control (QC) testing before, during and/or after cell expansion, such as tests for identity, potency, purity, and sterility, in accordance with GMP requirements. It should be understood that many QC assays may be conducted by the system, including without limitation cell-based assays, fluorescent-, colorimetric- or luminescent- based assays, cell morphology and cell time-dependent behavior (such as differentiation) assays, flow cytometry based assays, PCR based assays, endotoxin, mycoplasma and sterility assays, cell viability, cell number, cell confluency, and the like.

[0024] In some implementations, systems and methods provided herein include functionalities which expand cells and purify cells after expansion. In some implementations, systems and methods provided herein include functionalities which expand multiple cell lines at the same time without cross-contamination between cell lines. For example, functionalities may be included which ensure that no more than one sample is open at the same time in the enclosure. Similarly, reagent and supply containers are not opened when a sample container is open. Other included functionalities include those which reduce particle generation; allow sterilization of the system between cell processing steps; and functionalities for capping, uncapping, and recapping containers, which ensure that containers are not kept open longer than necessary and that containers are not open when or if a sample container is open; and the like. Particle monitoring can be used to pause processing steps until particle counts have gone below a pre-set threshold that ensures no cross-contamination between samples, and/or no cross -contamination from samples to stock reagents. Such functionalities facilitate processing of multiple batches at the same time without cross-contamination between batches.

[0025] In some implementations, systems and methods provided herein include functionalities that isolate cells from a starting tissue sample in preparation for further expansion or other processing.

[0026] In some implementations, systems and methods provided herein include functionalities that freeze or thaw cells.

[0027] In some implementations, systems and methods provided herein include functionalities that package cells, e.g., for transport or storage.

[0028] In some implementations, systems and methods provided herein include functionalities that provide cells in vials or cassettes for transport or storage.

[0029] In some implementations, systems and methods provided herein include one or more, two or more, three or more, or all of the following functionalities: 1) isolation of cells from starting tissue or from a mixture of various cell types; 2) identification and tracking of cell samples, e.g., using barcodes, positional information, and the like; 3) cell processing, e.g., expansion, purification (including enrichment or depletion, e.g. via magnetic antibodies), activation, reprogramming, gene editing (gene insertion, deletion, correction), transfection, and other desired manipulations of cells. Functionalities for analytical, e.g., marker expression level analysis (e.g., via fluorescent antibody staining and analysis), cell behaviour analysis including determination of differentiation profile, diagnostic testing to identify e.g. gene mutations, and QC testing including tests for identity, purity and sterility (optionally including endotoxin and mycoplasma testing), as well as for determination of cell number, confluency and viability, may also be included and can be conducted at any time before, during or after cell processing; 4) storage and transport, e.g., freezing cells in vials if desired or placing live cultures in a transport container (such as a Petaka™ cassette), packaging cells for transport, and the like; and 5) additional cell analytical capabilities as desired, such as purification of desired cell types, selection of a desired potency, removal of dead cells, magnetic cell sorting, and the like.

[0030] In some implementations, systems and methods provided herein include functionalities which provide a complete record of cell processing from start to finish for Quality Assurance (QA) verification, in accordance with GMP requirements. The system can verify that all steps were performed properly and check all assay results (e.g., pass/fail results). Further, systems and methods may include functionalities for tracking batches, e.g., using barcodes and positional memory, in accordance with GMP guidelines. Further, QA analysis may include testing for sterility, contaminants (such as endotoxin and mycoplasma), and other tests as may be desired in accordance with GMP guidelines and other applicable regulations.

[0031] In some implementations, systems and methods provided herein include one or more, two or more, three or more, or all the following functionalities: 1) cell processing; 2) quality control; 3) quality assurance; 4) harvesting of cells and preparation for storage or transport and 5) analytical testing of cells (such as, without limitation, diagnostic testing). In some implementations, systems and methods provided here may further include functionalities for sample preparation, e.g., for isolating cells for processing from a starting biological sample.

[0032] In some implementations, systems and methods provided herein include a functionality which handles reagents under GMP conditions. Reagents are automatically imported into the enclosure, verified (e.g., using a barcode reader), opened, dispensed into aliquots, and stored by the system. Such reagents can be automatically introduced into the enclosure in the manufacturer's packaging, obviating the need for a human to open a reagent container. In some implementations, a functionality which robotically transports materials into and out of the enclosure is included. In some implementations, a functional testing of a reagent is performed to ensure it meets specifications, optionally together with sterility, endotoxin and/or mycoplasma testing.

[0033] In some implementations, systems and methods provided herein include a control unit which performs fully automated processing without human intervention. The control unit not only executes processing steps but decides which steps to follow in order to produce a desired end product. For example, the control unit can determine which steps to perform depending on assay data obtained at various steps during the processing.

[0034] In some implementations, systems provided herein comprise a plurality of systems connected together. For example, a first system may be connected to a second system through a freezer or an incubator which is placed between the two systems and connected separately to each one. Alternatively, two enclosures may be connected to each other. It should be understood that a plurality of systems can be connected together in this way, either directly (enclosure-to-enclosure) or through a shared component such as a freezer, a refrigerator, an incubator, etc. The number of systems that can be connected in this way is not particularly limited.

[0035] In some implementations, systems provided herein comprise one or more, two or more, three or more, four or more, five or more, more than five, or all of the following automated components, or a combination thereof: (1) a robotic aspirator with disposable tips with the capability of changing the tip after each use or between samples, such that cross-contamination between samples is reduced or eliminated without requiring sterilization of the robotic aspirator component; (2) one or more decapper modules, for opening and closing a screwcap lid of containers, including large (>10 ml containers); (3) a centrifuge, cell sorter or magnet, e.g., for purifying cell mixtures (which can optionally also be achieved by e.g. magnetic cell separation) or obtaining a cell pellet or for collection or removal of cells; (4) an incubator for incubating cells; (5) a confluency reader or cell counter for determining cell number and/or cell confluency in a sample or in a cell-containing vessel; (6) a direct liquid to plate fill station or continuous flow robotic reagent dispenser for dispensing a volume of liquid directly into a cell-containing vessel (e.g., volumes > 5 ml); and (7) a tilt module for aspiration or collection of cells or of cell culture media, optionally as a magnetic separation tilt module.

[0036] In some implementations, systems provided herein comprise a sealed enclosure configured to minimize particle generation, e.g.: including a centrifuge placed below deck and sealed from the enclosure during use; including a vertical waste chute in which solid waste is dropped, sized so that waste does not hit the edges of the chute during disposal, and placed under strong enough negative pressure so no entry of particles from the chute into the enclosure occurs; including closable vents for sealing the enclosure to allow sterilization of the enclosure; including a functionality which provides rapid clean air for rapidly exchanging all the air in the system with clean air of the system; and other such functionalities and components as are described herein.

[0037] In some implementations, systems provided herein comprise a magnetic separation tilt module, e.g., for magnetic separation or transfection of cells. In some implementations, systems provided herein comprise an on-deck temperature-controlled freezer, such as a Grant freezer, for freezing of samples or to allow manipulation of samples and reagents at subzero temperatures.

[0038] In some implementations, systems provided herein comprise a tilt module configured to hold cell culture transport trays (such as Petaka™ trays) for loading or removing samples from transport trays.

[0039] In some implementations, systems provided herein comprise autoclavable bottle or tube holders that hold bottles or tubes to allow automated decapping and capping as well as automated transport of the bottle(s) or tube(s) within the system.

[0040] In some implementations, systems provided herein comprise a direct fill to cell processing container media fill station with dripping and overflow control.

[0041] In some implementations, systems provided herein comprise a robotic aspirator with changeable, sterile, disposable tips, with the capability of the system changing the tip by itself (without human intervention) after each use or between samples, such that cross-contamination between samples is eliminated or reduced without requiring sterilization of the vacuum aspirator component. In some implementations, the robotic aspirator further comprises an integrated tube and tip gripper. The robotic aspirator is designed to prevent any backflow or dripping by maintaining continuous negative pressure through the tip orifice (until disposal), and the tip being replaced between each use or batch. The fluid flow channels of the robotic aspirator through which aspirated fluid flows away from the tip can be further sterilized at, for example, the bleach station at regular intervals.

[0042] In some implementations, systems provided herein comprise autoclavable tip holders with system-closable lids, i.e., lids that can be opened and closed using robotic systems.

[0043] In some implementations, systems provided herein comprise a robotic module for robotic transport of materials into and out of the enclosure.

[0044] In some implementations, systems provided herein comprise a module for collecting biologicals and other macromolecules secreted or produced by cells, which can be optionally further purified and/or tested for identity, potency (e.g., activity assays) and/or sterility, and optionally vialed and/or freeze-dried and/or packaged.

[0045] In some implementations, systems and methods provided herein are fully automated, the above functionalities being carried out without human or hands-on intervention.

[0046] In some implementations, the fully automated systems and methods provided herein are conducted in a fully-enclosed processing environment that is aseptic and able to meet regulatory requirements for a "clean room", e.g., GMP requirements, CLIA requirements, and the like. Further, a plurality of batches can be processed at the same time under these conditions without cross-contamination between batches. In another broad aspect, there are provided methods for processing biological samples using the automated systems and methods described herein.

[0047] In another broad aspect, there are provided batches and biological samples prepared using the automated systems and methods described herein. A wide variety of biological materials may be prepared using the systems and methods described herein, including without limitation cells, tissue matrices, proteins, antibodies, vaccines, therapeutics, extracellular matrix components, and the like. In some implementations, cells are stem cells, stem-like cells, unipotent cells, multipotent cells, pluripotent cells, somatic cells, cell lines, immortalized cells, yeast or bacterial cells. Such cells may be prepared for example through reprogramming, transformation, or differentiation from another cell type. In particular implementations, the cells are autologous cells that are prepared from a starting biological sample from a patient for transplantation back into the same patient, e.g., autologous stem, stem-like, multipotent, unipotent, or somatic cells prepared for therapeutic use in the patient. In some implementations, the cells prepared are neural stem cells, neural stem-like cells, neural precursor cells, neural progenitor cells, neuroblasts, neurons, cardiac cells, hematopoietic cells, cells of ectoderm, mesoderm or endoderm lineage, pluripotent cells, multipotent cells, unipotent cells, somatic cells, naturally occurring cells, non-naturally occurring cells, prokaryotic cells, and/or eukaryotic cells. It should be understood that many different types of cells may be prepared using systems and methods described herein, and the type of cell is not meant to be limited.

[0048] In one implementation, there is provided a unipotent or multipotent cell prepared using the automated systems and methods described herein. In another implementation, there is provided a population of multipotent, unipotent, somatic, or stem-like cells prepared using the automated systems and methods described herein.

[0049] In some implementations, there are provided methods for reprogramming a cell of a first type to a desired cell of a different type that is multipotent or unipotent using the automated systems and methods described herein, the cell of a first type being a somatic cell, a stem cell, or a progenitor cell, the automated process executable by the systems described herein, the methods comprising steps of: introducing into the cell of a first type using robotic means an agent capable of remodeling the chromatin and/or DNA of the cell, wherein the agent capable of remodeling the chromatin and/or DNA is a histone acetylator, an inhibitor of histone deacetylation, a DNA demethylator, and/or a chemical inhibitor of DNA methylation; transiently increasing intracellular levels of at least one reprogramming agent in the cell of a first type using robotic means, wherein the at least one reprogramming agent increases directly or indirectly the endogenous expression of at least one multipotent or unipotent gene regulator to a level at which the gene regulator is capable of driving transformation of the cell of a first type into the multipotent or unipotent cell; using robotic means to maintain the cell of a first type in culture conditions supporting the transformation of the cell of a first type to the multipotent or unipotent cell for a sufficient period of time to allow a stable expression of a plurality of secondary genes characteristic of the phenotypical and/or functional properties of the multipotent or unipotent cell, where one or more of the secondary genes is not characteristic of phenotypical and functional properties of an embryonic stem cell and wherein stable expression of the plurality of secondary genes occurs in the absence of the reprogramming agent, whereby at the end of said period of time the cell of a first type has been transformed into the multipotent or unipotent cell, and where the multipotent or unipotent cell expresses at least one marker characteristic of the cell of a first type.

[0050] In another broad aspect, there is provided a robotic aspirator comprising: a robotic arm configured to move in at least one direction; a body connected to the robotic arm; and an aspiration member comprising a fluid flow channel connected to the body, the aspiration member being configured for connection to a pump means; the body being configured to hold a disposable tip for providing fluid connection between the disposable tip and the fluid flow channel of the aspiration member; fluid being aspirated through the disposable tip and the fluid flow channel when the disposable tip is fluidly connected to the fluid flow channel and the aspiration member is connected to the pump means. In some implementations, the robotic aspirator further comprises a plurality of prongs connected to the body, the prongs being moveable between a tip holding position and a retracted position, the prongs being configured in the tip holding position to hold the disposable tip for providing fluid connection between the disposable tip and the fluid flow channel of the aspiration member. In some implementations of the robotic aspirator, the disposable tips are capable of being disengaged from the fluid flow channel without handling by a human operator. In some implementations the prongs can hold tubes.

[0051] Methods of automatically aspirating a sample using the robotic aspirator described herein are also provided. In some implementations, there is provided a method of aspirating using a robotic arm having a fluid flow channel and a plurality of prongs configured to selectively hold a disposable tip in fluid connection with the fluid flow channel, the method comprising: moving the prongs to retain the disposable tip in fluid connection with the fluid flow channel, the prongs being selectively moveable and optionally further configured to grip at least one object other than the disposable tip; and evacuating the fluid flow channel to aspirate liquid through the disposable tip and the fluid flow channel. In some implementations, the method comprises, after aspirating liquid, disengaging the prongs from the disposable tip; and stopping evacuation of the fluid flow channel to disengage the disposable tip from the fluid flow channel. In some implementations, the disposable tip disengages from the fluid flow channel without handling by a human operator.

[0052] Embodiments of the present invention each have at least one of the above-mentioned object and/or aspects, but do not necessarily have all of them. It should be understood that some aspects of the present invention that have resulted from attempting to attain the above-mentioned object may not satisfy these objects and/or may satisfy other objects not specifically recited herein.

[0053] Additional and/or alternative features, aspects, and advantages of embodiments of the present invention will become apparent from the following description, the accompanying drawings, and the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0055] For a better understanding of the present invention, as well as other aspects and further features thereof, reference is made to the following description which is to be used in conjunction with the accompanying drawings, where:

[0056] Figure 1A is a perspective view, taken from a front, top and left side, of an automated cell processing system (ACPS) in accordance with an implementation of the present technology;

[0057] Figure IB is a front elevation view of the ACPS of Figure 1 A;

[0058] Figure 1C is a top plan view of the ACPS of Figure 1 A;

[0059] Figure 2 is a schematic illustration of the ACPS of Figure 1 A;

[0060] Figure 3 A is a perspective view, taken from a front and right side, of a portion of the ACPS of Figure 1A with an isolator, a biological safety cabinet and a control unit removed for clarity;

[0061] Figure 3B is a perspective view, taken from a front and right side, of the portion of the ACPS of Figure 3A, with the top wall and side walls of the enclosure being removed for clarity;

[0062] Figure 3C is a perspective view, taken from a front and right side, of the bottom wall of the enclosure and the table of Figure 3A shown in isolation;

[0063] Figure 4 is a top plan view of components housed within the enclosure, the bottom wall of the enclosure and the isolator of Figure 3 A;

[0064] Figure 5 is a close up perspective view, taken from a front and right side, of some of the components housed within the enclosure of Figure 4;

[0065] Figure 6 is a perspective view, taken from a front and right side, of a left rack of a storage area of Figure 5;

[0066] Figure 7 is a perspective view, taken from a front and right side, of a right rack of a storage area of Figure 5;

[0067] Figure 8A is a perspective view, taken from a front and right side, of a center portion of a storage area of Figure 5;

[0068] Figure 8B is a close up perspective view, taken from a front, top and left side, of one of the transfer trays of the center portion of the storage area of Figure 8B;

[0069] Figure 9 is a top plan view of a deck housed within the enclosure of Figure 3A;

[0070] Figure 10A is a perspective view, taken from a front, top and left side, of a portion of the deck 910 of Figure 9 showing a media fill station and a magnetic tilt module;

[0071] Figure 10AB is a close-up perspective view, taken from a front, top and left side, of the magnetic tilt module of Figure 10A;

[0072] Figure IOC is a perspective view of another implementation of a media fill station with a dispensing tip in a load position;

[0073] Figure 10D is a perspective view of the media fill station of Fig. IOC with the dispensing tip in a fill position;

[0074] Figure 11A is a perspective view, taken from a rear, top and left side, of a portion of the deck 910 of Figure 9 showing a tilt module for cell processing trays, a media fill station, transport container tilt module, and a transport holder adaptor station;

[0075] Figure 1 IB is a close-up exploded perspective view, taken from a front and right side, of the transport container tilt module;

[0076] Figure 11C is a close-up perspective view, taken from a front and right side, of the transport container tilt module shown in an untilted position;

[0077] Figure 1 ID is a close-up perspective view, taken from a front and right side, of the transport container tilt module shown in an untilted position;

[0078] Figure 12 is a perspective view, taken from a left, rear and top, of a portion of the ACPS of Figure 3A with the upper wall and side walls of the enclosure being removed for clarity and showing the waste receptacle;

[0079] Figure 13 is a perspective view, taken from a front and top of an example robotic arm of a robotic module of the ACPS of Figure 3A having a robotic aspirator/gripper;

[0080] Figure 14 is a front plan view of the example robotic aspirator/gripper of Figure 14 holding a tube;

[0081] Figure 15 is a front plan view of the example robotic arm of Figure 14 having an aspirator tip attached thereto;

[0082] Figure 16 is a perspective view, taken from a front, top and left side, of another robotic arm having a decapper and shown decapping a reagent container;

[0083] Figure 17 is a perspective view, taken from a front, top and right side a first ACPS connected to a second ACPS;

[0084] Figure 18 is a schematic view of the control unit of the ACPS 100 for executing a method in accordance with an implementation of the present technology;

[0085] Figure 19 is a schematic illustration of an example robotic module of the ACPS of Figure 4;

[0086] Figure 20A is a perspective view, taken from a rear, top and right side of, a flask used in the ACPS of Figure 3A;

[0087] Figure 20B is a right side elevation view of the flask of Figure 20A placed on a tilt module with the tilt module disposed in an untilted position;

[0088] Figure 20C is a front elevation view of the multilayer flask and multilayer flask tilt module of Figure 20B with the tilt module being disposed in a tilt position with the multilayer flask being tilted about a longitudinal tilt axis;

[0089] Figure 20D is a right side elevation view of the flask of Figure 20A placed on the multilayer flask tilt module of Figure 20B with the multilayer flask tilt module being tilted about a lateral tilt axis;

[0090] Figure 21 is a top plan view of another implementation of a deck housed within the enclosure of Figure 3A;

[0091] Figure 22 is a close-up perspective view, taken from a bottom, front and right side of two air outlets of the enclosure of Figure 3A along with the corresponding automated gates for selectively closing the air outlets;

[0092] Figure 23 is a close-up perspective view, taken from a bottom, front and right side, of one of the air outlets and corresponding gate of Figure 22 with the gate being shown in a position where the air outlet is fully closed;

[0093] Figure 24 is a close-up perspective view, taken from a bottom, front and right side, of the air outlet and gate of Figure 23 with the gate being shown in a position where the air outlet is fully open;

[0094] Figure 25 is a rear elevation view of a portion of the ACPS of Figure

3 A showing another implementation of a waste receptacle and waste chutes;

[0095] Figure 26 is a perspective view, taken from a top, front and left side, of the waste receptacle of Figure 25 shown in isolation;

[0096] Figure 27 is a cross-sectional view, taken along a plane extending vertically and laterally through the ACPS and the waste receptacle and waste chutes of Figure 25; and

[0097] Figure 28 is a schematic illustration of the automated method 2000 for cell processing. Figure 29A is a close-up perspective view of a transport tray according to some implementations;

[0098] Figure 29B is a close-up perspective view of a cell processing tray according to some implementations;

[0099] Figure 29C is a close-up perspective view of a storage tube according to some implementations;

[00100] Figure 29D is a close-up perspective view of a centrifuge tube according to some implementations;

[00101] Figure 30A is a perspective view, taken from a front, top and right side, of a holder for the storage tubes of Figure 29C showing one of the storage tubes in the holder being gripped by the robotic aspirator/gripper of Figure 14; and

[00102] Figure 30B is a perspective view, taken from a front, bottom and right side of the holder of Figure 30A;

[00103] Figure 31A is a top plan view of another implementation of a cell processing tray;

[00104] Figure 3 IB is a front elevation view of the cell processing tray of

Figure 31 A;

[00105] Figure 31C is a side elevation view of the cell processing tray of Figure 31 A; and

[00106] Figure 3 ID is a cross-sectional view of the cell processing tray of

Figure 31 A, taken along the line 3 ID.

DETAILED DESCRIPTION

[00107] There are described herein methods and systems that can be used for transforming a cell of a first type, such as a somatic cell, a stem cell, or a progenitor cell, to a cell of a desired second type, such as a pluripotent, multipotent, or unipotent cell. The described methods and systems are provided in order to illustrate certain implementations of the methods and systems. It should be expressly understood that other implementations are possible. In particular, it should be understood that methods and systems can be used for a wide variety of biological sample processing, including generation of biomaterials (e.g., tissues, matrices, etc.), generation of biologies (e.g., proteins, antibodies, growth factors, etc.), growth of cells and cell lines, in addition to cell transformation and cell reprogramming.

[00108] With reference to Figs. 1A to 2, an automated cell processing system

(ACPS) 100 for an automated method of cell processing includes an enclosure 110. The enclosure 110 is connected to an isolator 120 and via the isolator 120 to a biological safety cabinet (BSC) 130.

[00109] The ACPS 100 also includes various equipment such as refrigerators, incubators, freezers and the like some of which are disposed inside the enclosure 110, the isolator 120, or the BSC 130, and some of which are disposed outside the enclosure 110, the isolator 120, and/or the BSC 130, so as to be accessible from within the enclosure 110, the isolator 120, and/or the BSC 130.

[00110] The ACPS 100 includes a control unit 1000 configured to control the automated cell processing as will be described in further detail below.

Enclosure

[00111] With reference to Figures 1A to 3A, the enclosure 110 is a rectangular chamber constructed of four side walls 202, 204, 206, 208, an upper wall 210, and a bottom wall 212. The side walls include a front wall 202, a rear wall 204, a left side wall 206 and a right side wall 208. Terms such as left, right, front and rear are defined herein as would be understood by a person standing on the bottom wall 212 within the enclosure 110 and facing forwardly towards the isolator 120. The walls are made of metal but it is contemplated that the walls could be made of any suitable material.

[00112] The front wall 202 has an isolator connection port 220 which connects to a complementary port 240 of the isolator 120. The isolator connection port 220 is rectangular in shape but it is contemplated that the isolator connection port 220 could be other than rectangular. The isolator connection port 220 is normally closed by a gate (not shown) and opened only to allow transfer objects between the enclosure 110 and the isolator 120. The enclosure 110 is thus in selective fluid connection with the isolator 120.

[00113] Eight air inlets 222 are defined in the upper wall 210 of the enclosure.

Each air inlet 22 has a HEPA (High Efficiency Particulate Air) or ULPA (Ultra Low Particulate Air) filter (not shown). An air flow system which includes impellers mounted inside the enclosure 110 pushes air into the enclosure 110 through the HEPA filter provided in the air inlet port 222 and maintains circulation of air through the enclosure 110. It is contemplated that there could be more than one air inlet 222. It is contemplated that other appropriate air filter, such as an ULPA (Ultra Low Penetration Air) filter, could also be used in place of the HEPA air filter.

[00114] Two air outlets 224 are formed in the bottom wall 212. Additional air outlets 225 (Fig. 3A) are also provided near the bottom of the front wall 202 and the bottom of the rear wall 204. It is contemplated the number and configuration of the air outlets 224 could be different than as shown. In some implementations, airflow in the enclosure 110 is laminar. In some implementations the laminar airflow can be used to divide the space within the enclosure 110 into a plurality of portions. The portions inside the enclosure 110 created by the laminar flow could be used to process different batches, as will be described below in further detail, without increasing risk of cross contamination between the batches. The enclosure 110 is maintained at a positive air pressure relative to the ambient pressure in the room housing the automated cell processing system 100, and relative to the isolator 120. Rapid air exchange in the enclosure 110 helps to remove any contaminant particles that may have entered the enclosure 110 and thereby reduces the probability of exposure of objects housed inside the enclosure 110 to the contaminants that enter the enclosure 110.

[00115] The air outlets 224 along the floor 224 are closeable (for example, during sterilization of the enclosure 110) by automated gates 250. The air outlets 225 formed defined in the front and rear walls 202, 204 are also closeable (for example, during sterilization of the enclosure 110). All of the outlets 224 formed in the bottom wall 212 are generally similar and as such, one of the outlets 224 and the automated gate 250 covering the outlet 224 will now be described. With reference to Figs. 22 to 24, the outlet 224 is covered with a mesh screen 251 which is made of stainless steel in the illustrated implementation. It is contemplated that the screen 251 could be made of any suitable material. The screen 251 ensures and prevents objects from outside the enclosure 110 from entering the inside of the enclosure 110, or objects inside the enclosure 110 from falling through the outlet 224. The gate 250 is slidably mounted to a pair of flanges 252 mounted on opposite sides of the outlet 224. The flanges 252 are generally mirror images, each having a groove 253 facing the groove 253 of the opposite flange 252. The opposing grooves 253 extend parallel to the bottom wall 212 except at the end where each groove 253 forms a ramp 255 bending towards the bottom wall 212. The gate 250 has two guiding elements 254 connected along each side, one at each end of the side of the gate 250. Each guiding element 254 is shaped and sized to be received in the groove 253 and slide or roll therein. The guiding elements 254 move along the grooves 253 to guide the gate 250 between a closed position where the outlet 224 is sealed and an open position. In the closed position, one of the guiding elements 254 on each side is received in the ramp 255 bending towards the bottom wall 212. The ramp 255 pushes the gate 250 towards the bottom wall 212 to ensure sealing between the gate 250 and the bottom wall 212. In the open position, the guiding elements 254 are disposed in groove 253 outside the groove end 255. An electrical actuator 256 is connected to the gate 250 for moving the gate 250 so as to slide or roll the guiding elements 254 along the corresponding grooves 253. The actuator 256 is connected to the control unit 1000 for controlling the opening and closing of the air outlets 224. In the illustrated implementation, the actuator 256 is controlled to move the gate 250 between a position where the air outlet 224 is fully open or a position where the air outlet 224 is fully closed. It is contemplated that the gate 250 could be controlled to maintain the gate 250 in a position where the outlet 224 is partially open.

[00116] A sterilant inlet 230 is defined in the left side wall 206 for introducing sterilant into the enclosure 110 for sterilization of the space inside the enclosure 110. The sterilant inlet 230 is configured for attachment of a fluid conduit to receive sterilant (in gas or vapour form in the illustrated implementation) and to deliver the received sterilant into the interior of the enclosure 110 as a sterilant vapor mist or spray. The sterilant air inlet 230 has a cover to prevent entry of foreign particles when not in use.

[00117] A sterilant outlet 232 is also defined in the front wall 202 for removing air and sterilant from the enclosure 110. The sterilant outlet 232 is configured for attachment of a fluid conduit leading to a pump for removing sterilant vapour, gas or air from the enclosure 110.

[00118] A catalytic converter inlet 231 is defined in the left side wall 206 for introducing air into the enclosure for recirculating air through a catalytic converter to convert the sterilant vapor to harmless and biodegradable water vapor and oxygen at the end of a sterilization procedure. The catalytic converter inlet 231 is configured for attachment of a fluid conduit and has a cover to prevent entry of foreign particles when not in use.

[00119] A catalytic converter outlet 233 is also defined in the left side wall

206 above the HEPA or ULPA filters and configured for removing air from the enclosure through these HEPA and ULPA filters and through a catalytic converter in order to more rapidly neutralize the vapour sterilant otherwise lodged into the extensive surface area of the HEPA or ULPA filters. The catalytic converter outlet 233 is configured for attachment of a fluid conduit leading to a catalytic converter and a pump for removing air and sterilant vapour from the enclosure 110.

[00120] It is contemplated that the sterilant inlet and outlet 230, 232 could each be defined in a location other than that shown herein, and configured differently than as shown herein. It is contemplated that the catalytic converter inlet and outlet 231, 233 could each be defined in a location other than that shown herein, and configured differently than as shown herein.

[00121] The sterilant inlet and outlet 230, 232 are connected to an automated enclosure sterilization unit 550 for decontamination of the interior of the enclosure 110. The automated enclosure sterilization unit 550 will be described below in further detail.

[00122] Various access ports are provided in the walls of the enclosure 110. In the bottom wall 212, as can be seen best in Fig. 3C, the enclosure 110 has access ports 170, 172, 174, 176, 178 for accessing various process equipment such as a centrifuge 150, a freezer 152, an incubator 154, and a waste receptacle 156. The bottom wall 212 also defines recesses 171 and 175 in which a robotic module 600 and a cryofreezer 460 are mounted respectively. It is contemplated that one or both of the recesses 171, 175 could be omitted or that other recesses could be formed for mounting of other components. An access port defined in the left side wall 206 is closed by a side panel 184.

[00123] It should also be understood that the number, shape, size, position and configuration of the ports of the enclosure 110 could be other than that shown herein. It should also be understood that the number, shape, size, position and configuration of the inlets and outlets (such as for air, sterilant and the like) of the enclosure 110 could be other than that shown herein.

[00124] The enclosure 110 generally remains sealed except for transferring objects (samples, reagent containers, containers for samples, other labware, and the like) between the enclosure 110 and the isolator 120, or other process equipment, such as incubators, centrifuges, freezers, storage cabinets and the like that may be connected to the enclosure 110 for the automated processing of cells. The connection between the enclosure 110 and these other process equipment is a sealed connection, and the enclosure 110 is maintained at a positive pressure relative to the interior of the process equipment to reduce entry of contaminant particles from the process equipment into the enclosure 110.

[00125] The enclosure 110 is generally considered a sterile / aseptic environment and maintained as a class 10 cleanroom (having fewer than 10 particles of a size greater than or equal to 0.5 microns per cubic square foot) in order to conform with good manufacturing practice (GMP) guidelines. The terms "sterile" and "aseptic" are used interchangeably herein to mean microbially sterile, i.e., not contaminated by microorganisms such as endotoxin, mycoplasma, bacteria, etc., or by

other infectious agents such as viruses. Thus it should be understood that the enclosure 110 is designed to be aseptic and microbial-free and this is determined by assays and processes in the system that test and measure for microbial contamination, such as endotoxin, mycoplasma, and direct microbial detection assays, to ensure that samples/batches are not contaminated.

[00126] The term "good manufacturing practice (GMP)" is used to refer to regulations for medicinal products established by government regulatory bodies such as the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMEA) to ensure safety and efficacy of products for clinical use. As used herein, the term "under GMP conditions" means under conditions that meet Good Manufacturing Practices (GMP) guidelines or regulations, i.e., so that the end product can be released for clinical use. It is noted that GMP regulations and recommended guidelines may vary nationally but in general require strict control in GMP production facilities for the manufacturing of pharmaceutical or cellular products, including quality control and quality assurance programs. Such facilities typically require "clean rooms", which are classified in four classes (AD) depending on air purity, based on the number of particles of two sizes (> 0.5 μιη, > 5 μιη), or are in accordance with Clinical Laboratory Improvement Amendments (CLIA) regulations; other parameters such as temperature, humidity, and pressure are often taken into account and monitored because of their potential impact on particle generation and microorganism proliferation; materials and staff flows are separated and unidirectional to minimize cross contamination; documentation of all activities is necessary; and so on. GMP regulations for cell therapy products generally include at least some of the following: demonstration of preclinical safety and efficacy; no risk for donors of transmission of infectious or genetic diseases; no risk for recipients of contamination or other adverse effects of cells or sample processing; specific and detailed determination of the type of cells forming the product and what are their exact purity and potency; and in vivo safety and efficacy of the product.

[00127] As can be seen best in Fig. 3C, the enclosure 110 is supported on a rectangular frame 140 having an upper portion 142 formed by upper horizontal frame members and a lower portion 144 formed by lower horizontal frame members. The frame 140 includes vertical frame members 143 extending between the upper and

lower horizontal frame members 142, 144. The lower portion 144 is supported on wheels to facilitate repositioning of the frame 140 but it is contemplated that the wheels could be omitted. The lower portion 144 supports other components of the ACPS 100 as will be described below. In some implementations, such as that shown in Figs. 1A to 1C, upper portion 142 is supported on the floor by the vertical frame members 143 with the lower portion 144 and the wheels being omitted.

[00128] The centrifuge 150, the incubator 152, freezer 154, and the waste receptacle 156 are supported on the lower portion 144. The centrifuge 150 has an access port on its upper portion, and is supported on the lower portion 144 such that the access port (not labeled) of the centrifuge 150 is aligned with the corresponding centrifuge access port 170 of the lower bottom wall 212. The space inside the centrifuge 150 is thus accessible from inside the enclosure 110 via the aligned access ports in the bottom wall 212 of the enclosure 110 and the upper portion of the centrifuge 150. Similarly, each of the incubator 152, freezer 154, and the waste receptacle 156 has an access port defined in their respective upper walls. The incubators 152, freezer 154, and the waste receptacle 156 are each supported on the lower portion 144 so as to align their respective access ports with the corresponding access port of the bottom wall 212 of the enclosure 110. It is contemplated that the lower portion 144 could be omitted and one or more of the centrifuge 150, the incubators 152, freezer 154, and the waste receptacle 156 could be placed on the room floor below the upper portion 142 supporting the enclosure 110. It is also contemplated that the one or more of the centrifuge 150, the incubator 152, freezer 154, and the waste receptacle 156 could be connected to a wall of the enclosure 110 other than the bottom wall 212. For example, the side walls of the enclosure 110 could have access ports (such as the access port covered by side panel 184) for connecting to one or more of the centrifuge 150, the incubator 152, freezer 154, and the waste receptacle 156.

[00129] A panel 226 mounted on the wall of the BSC 130 includes a display for pressure and other environmental characteristics of the enclosure 110 and manual override switches for various elements inside the enclosure 110 such as a light switch, impellers associated with air inlets 222, other mixing fans used during sterilization of the enclosure 110, and the like, which are controlled automatically by the control unit 1000 during routine operation of the ACPS 100.

[00130] The enclosure 110 houses various components of the ACPS 100 as will be described below.

Isolator

[00131] With reference to Figures 1A to 2, the isolator 120 is disposed in front of the front side wall 202 of the enclosure 110. The isolator 120 is a generally rectangular chamber defined by four side walls, an upper wall and a lower wall. The rear side wall has an enclosure access port 240 connected to the isolator access port 220 of the enclosure 110. A gasket (not shown) is installed around the enclosure access port 240 for forming a sealed connection between the isolator 120 and the enclosure 110. The enclosure access port 220 and the isolator access port 220 are selectively covered by a gate that is opened for passing objects (such as chemical supplies, lab ware, tissue samples, and the like) between the isolator 120 and the enclosure 110. The gate is an automated gate that is connected to the control unit 1000 for controlling the opening and closing of the ports 220, 240 connecting the isolator 120 to the enclosure 110.

[00132] The front wall of the isolator 120 is in the form of a hinged window

243 (hingedly connected at the upper edge in the illustrated implementation) and can be opened to access the interior space of the isolator 120 for cleaning and maintenance, for example. In the illustrated implementation, the front wall 243 is made of tempered glass but it could be made of any suitable material. It is contemplated that the front wall could be fixed and not openable for access to the interior. Four glove ports 242 (the gloves being removed in the figures for clarity) are provided in the front wall 243 to allow a human user to manipulate objects placed inside the isolator 120 while maintaining the environmental isolation and sterility of the interior of the isolator 120. In the illustrated implementation, the passage of objects between the isolator 120 and the enclosure 110 occurs via the automated transfer trays 322 (Fig. 3B). When the transfer tray 322 is extended into the isolator 120 through the ports 220, 240, a human operator using the glove ports 242 moves objects between the transfer tray 322 and the isolator 120. It is contemplated that a robotic module could be provided in the isolator for moving objects between the transfer tray 322 and the isolator 120 and/or the BSC 130. It is also contemplated that the transfer trays 322 could be manually actuated instead of or in addition to being electrically actuated. It is also contemplated that the passage of objects between the isolator 120 and the enclosure 110 could be performed fully manually, i.e. by a human operator using the glove ports 242 to transfer objects through the ports 220, 240 with or without the use transfer trays 322.

[00133] A BSC connection port 244 is defined in the right side wall of the isolator 120 for connection to the BSC 130. A sealed door (not shown) extending across the port 244 can be opened to allow passage of objects between the BSC 130 and the isolator 120. An interlock mechanism is provided to ensure that the enclosure access port 240 is closed when the BSC connection port 244 is open and vice versa.

[00134] The isolator 120 has two air inlets 246 provided with a HEPA air filter and an air outlet 248 for maintaining circulation of HEPA filtered air through the isolator 120. A sterilant outlet 234 is also provided top wall of the isolator for removing air and sterilant from the isolator 120. The isolator 120 can thus be sterilized via a sterilization unit (for example the sterilization unit 550) connected to the enclosure 110 by keeping the enclosure connection port 240 open during sterilization of the enclosure 110. The sterilant outlet 234 is configured for attachment of a fluid conduit leading to a pump for removing sterilant vapour, gas or air from the isolator 120. Impeller fans (not shown) are also provided in the isolator 120 to maintain optimal circulation of air and/or sterilant through the isolator 120. The isolator 120 is maintained at a positive air pressure relative to the BSC 130 and at a negative pressure relative to the enclosure 110 so that air flows out of the enclosure 110 into the isolator 120 when the connection ports 220, 240 are open, thereby reducing the possibility of contamination due particles entering the enclosure 110 from outside. It is contemplated the number and configuration of the air inlets and outlets 246, 248 could be different than as shown herein. The isolator has an access port 245 (shown schematically in Fig. 2) on the right side wall for connection to a refrigerator 160 for storing reagent and other media containers.

[00135] The isolator 120 is used to transfer samples and other objects from larger containers to smaller containers before passing into the enclosure 110. In some implementations, the outer protective packaging of objects may be removed in the isolator 120 before passing into the enclosure 110. In some implementations, the isolator 120 could house one or more reagent containers.

[00136] In some implementations, the isolator 120 has an automated sterilization system (such as the system 550 shown schematically in Fig. 2) for sterilizing the isolator 120, for example with hydrogen peroxide.

Biological Safety Cabinet (BSC)

[00137] With reference to Figures 1A to 2, the BSC 130, also in the form of a generally rectangular chamber defined by four side walls, an upper wall and a lower wall, is disposed on a right side of the isolator 120. The BSC 130 has an isolator connection port 260 defined in its left side wall and connected to the BSC connection port 244 of the isolator 120. An access port 262 in the front wall of the BSC 130 is used for transferring objects into and out of the ACPS 100 by a human and/or robotic operator. The access port 262 is covered by a sliding gate 263 that is opened for transferring objects therethrough. In the illustrated implementation, the sliding gate 263 is made of tempered glass but it could be made of any suitable material. An interlock mechanism is provided to ensure that the isolator connection port 260 is closed when the access port 262 is open and vice versa. As shown schematically in Fig. 2, a transfer tray 132 mounted on rails 134 is used to transfer objects between the isolator 120 and the BSC 130. In the illustrated implementation objects can be placed on the transfer tray 132 by a human operator and the transfer tray 132 could be actuated manually to move the transfer tray 132 between the isolator 120 and the BSC 130. It is however contemplated that the transfer tray 132 could be electrically actuated and that objects could be moved to/from the transfer tray 132 robotically by a robotic arm provided in the BSC 130 and/or in the isolator 120.

[00138] The BSC 130 has an air inlet 266 covered with a HEP A air filter and an air outlet 268 for maintaining circulation of HEP A filtered air through the BSC 130. It is contemplated the number and configuration of the air inlets and outlets 266, 268 could be different than as shown herein. Impeller fans can be optionally provided in the BSC 130 to maintain air circulation through the BSC 130. The BSC 130 is maintained at a positive air pressure relative to the ambient air in the room housing the system 100, and at a negative pressure relative to the isolator 120 so that air flows out of the isolator 120 into the BSC 130 when the connection ports 244, 260 are open, thereby reducing the possibility of contamination due particles entering the enclosure 110 from outside. In the illustrated implementation, the BSC 130 is maintained as a class 100 cleanroom environment (having fewer than 100 particles of a size greater than or equal to 0.5 microns per cubic square foot). It is however contemplated that the BSC 130 could be maintained at a higher or lower level of cleanroom environment.

[00139] The BSC 130 is used to as a location to manually clean or sterilize the outer surface of objects (or the outer packaging of a container of sterile objects) before passing the objects into the isolator 120, and thereby into the enclosure 130. After sterilizing the outer surface of objects placed inside the BSC 130, the sliding gate is closed to cover the front access port 262. HEPA filtered air is then circulated through the BSC 130 for a predetermined amount of time to reduce the number of particles in the air before opening the isolator connection port 260 for passing objects from the BSC into the isolator 120.

[00140] It is contemplated that the configuration of any of the enclosure 110, the isolator 120, and the BSC 130 and/or the connections therebetween could be different than as shown herein. For example, the number, dimension, placement of the access ports in any one or more of the enclosure 110, the isolator 120, and the BSC 130 could be different. It is also contemplated that one or both of the isolator 120 and the BSC 130 could be omitted, for example if the enclosure 110 were placed in a cleanroom. It is further contemplated that isolator 120 and BSC 130 can be replaced by a robotic system that places sterile or aseptic materials on the tray 322 (or on another transport system) for introducing objects to or retrieving objects from enclosure 110.

[00141] If all the connecting ports 220, 240, 244, 260, 262 connecting between the enclosure 110 and the isolator 120, the isolator 120 and the BSC 130, and the BSC 130 and the external environment are open, air flows from the enclosure 110 to the isolator 120, from the isolator 120 to the BSC 130, and from the BSC to the room or external environment due to the positive pressure in the enclosure 110 relative to the isolator 120, the positive pressure in the isolator 120 relative to the BSC 130, the positive pressure in the BSC 130 relative to the room or external environment.

[00142] As mentioned above, in the ACPS 100, the enclosure 110 can access various equipment needed for the cell processing.

[00143] In the illustrated implementation of the ACPS 100, the centrifuge 150 is a Hettich™ Rotanta robotic centrifuge which includes a robotic arm inside the centrifuge for transferring objects into and out of the centrifuge 150. The centrifuge 150 is normally sealed from the enclosure 110 except for the sealed inner chamber of the centrifuge 150 being open to the space inside the enclosure 110 while samples are being loaded into and unloaded therefrom. The inner chamber of the centrifuge 150 is maintained at a slight negative pressure relative to the enclosure 110. The centrifuge 150 is installed under the deck 910 (described in further detail below) so that particles generated by the centrifuge 150 do not enter the enclosure 110 when the access ports 170 therebetween are open. The centrifuge 150 may be associated with a barcode reader or other device to verify and record the identity of containers entering and exiting the centrifuge 150 in order to track different steps during cell processing as desired for complying with GMP regulations. The control unit 1000 is communicatively coupled to the centrifuge 150 for automated cell processing.

[00144] In the illustrated implementation of the ACPS 100, the incubator 152 is a Liconic™ STR240 which includes a robotic arm inside the incubator for transferring objects into and out of the incubator 152. The incubator 152 is sealed from the enclosure 110 and maintained at a slight negative pressure relative to the enclosure 110 so that particles generated in the incubator 152 do not enter the enclosure 110 when the access ports 172 therebetween are open. In some implementations, the incubator 152 is constructed in a way that prevents contamination (for example, including features such as a chamber fully constructed of copper alloy, HEPA filters, a sterile water vapour generator instead of a water pan inside incubator, and the like). The incubator 152 is connected to an automated incubator sterilization unit 552 for decontamination of the interior of the incubator 152. The automated incubator sterilization unit 552 is disposed adjacent the incubator 152 and supported on the lower portion 144 of the frame 140. The automated incubator sterilization unit 552 will be described below in further detail. The

incubator 152 can be independently sterilized, for example using C102 gas, while the cells are in a secondary incubator or in the enclosure 110. The incubator 152 also has a barcode reader to verify and record the identity of containers entering and exiting the incubator 152 in order to track different steps during cell processing as desired for complying with GMP regulations. The control unit 1000 is communicatively coupled to the incubator 152 for automated cell processing and to the automated incubator sterilization unit 552 for sterilization of the incubator 152.

[00145] In the illustrated implementation of the ACPS 100, the freezer 154 is a

Liconic™ STR 44 which includes a lift 155 (Fig. 9) for transferring objects into and out of the freezer 154. The freezer 154 also has a barcode reader to verify and record the identity of containers entering and exiting the freezer 154 in order to track different steps during cell processing as desired for complying with GMP regulations. The control unit 1000 is communicatively coupled to the freezer 154 for automated cell processing. In the illustrated implementation, the freezer 154 is provided with a double door (one door 270 of the double doors being shown in Fig. 3C) instead of one door closeable to seal the enclosure 110 from the freezer 154. The door 270 is an insulation door for providing additional insulation and is automatically closed during sterilization of the enclosure 110 to prevent condensation of certain sterilants (hydrogen peroxide vapor, for example) around the freezer door which would be colder than the ambient temperature if the freezer door lacked the insulation door. The insulation door 270 is a slidable door mounted to the upper surface of the bottom wall 212 of the enclosure 110. The insulation door 270 is actuated by an electric actuator which connected to the control unit 1000 and thereby controlled by the control unit 1000 for closing of the insulation door 270 during sterilization procedures.

[00146] The refrigerator 160 in the illustrated implementation of the ACPS

100 is maintained at 4°C and used to store reagent containers. The interior of the refrigerator 160 is accessible via the isolator 120 through an access port in the right side of the isolator 120. The reagent container is placed in the refrigerator 160 by a human operator and connected to a media fill line which extends through the isolator 120 to a media fill station 420 in the enclosure 110. It is contemplated that the

refrigerator 160 could also be provided with a double door including an insulation door similar to the freezer 154 described above.

[00147] In some implementations, the ACPS 100 includes a robotic cr ostorage unit 162 (shown schematically in Fig. 2) for storing containers after cell processing has been completed. In the illustrated implementation, the cryostorage unit 162 is an Askion™ C-line System cryostorage unit. The cryostorage unit 162 is connected to the enclosure 110 by a sealed connection similar to that of the freezer 154 or centrifuge 150 as described above. The cryostorage unit 162 could also have its own robotic system (including for example a robotic arm) to allow automatically storing and retrieving of containers therefrom into the enclosure 110 without handling by a human operator.

Components of the ACPS Inside the Enclosure

[00148] With reference to Figs. 2, 3A, 3B and 4, inside the enclosure 110, the

ACPS 100 has a storage area 300, a sample preparation and processing area 400, a quality control area 500, a harvesting area 900 and robotic modules 600, 700 and 800, 820.

[00149] In the illustrated implementation of the ACPS 100, the storage area

300 is located proximate the front wall of the enclosure 110 rearward of the isolator connection port 220, and the robotic module 700 is disposed rearward of the storage area 300. In the illustrated implementation of the ACPS 100, the cell processing area 400 is located rearward of the robotic module 700, the robotic module 600 is disposed on a right side of the cell processing area 400 proximate the right side wall of the enclosure 110, and the robotic modules 800, 820 are disposed above the cell processing area 400. In the illustrated implementation of the ACPS 100, the harvesting area 900 is disposed on a left side of the cell processing area 400, and the quality control area 500 is disposed on a left side of the harvesting area 900. In some implementations, the quality control area 500 is also disposed vertically higher than the harvesting area 900 and the cell processing area 400.

[00150] Generally, the storage area 300 includes a plurality of storage modules, the processing area 400 includes a plurality of cell processing modules, the harvesting area 900 includes one or more harvesting modules and the quality control area 500 includes one or more quality control modules. Some modules may perform functions related to one or more of cell processing, harvesting and quality control, and thus these modules could be considered to be more than one type of module, for example, a cell processing module and a harvesting module. For example, a particular processing station, such as a tilt module could also be used for harvesting as will be described below. Additionally, any one or more of the areas (storage area 300, processing area 400, quality control area 500 and harvesting area 900) could be divided and located in physically separated locations. In the illustrated implementation in Fig. 2, the sample preparation and processing areas are shown in the same location, however they could be located in physically separated locations. Similarly, any combination of the above mentioned areas could be overlapping in the same location or could be located in physically separated locations.

[00151] In the illustrated implementation of the ACPS 100, the robotic module

700 accesses the storage area 300, the cell processing area 400, and the quality control area 500. In the illustrated implementation of the ACPS 100, the robotic module 600 accesses the right portion of the cell processing area 400 and the centrifuge 150. It is however contemplated that the relative position of the various components, areas and modules within the enclosure 110 could be different than as shown herein.

[00152] The ACPS 100 is configured for the robotic handling of various types of cell processing containers 314 including trays, flasks, bottles, tubes and vials. Examples of trays include cell processing trays 344 such as Omni™ trays shown in FIG. 29B, cell processing trays 344' shown in Fig. 31A to 3 ID, transport trays 340 such as Petaka™ trays shown in FIG. 29A, and the like. Examples of tubes include centrifuge tubes 346 (for example, Falcon™ tubes shown in Fig. 29D), storage tubes 884 (for example, Micronic TM tubes as shown in Fig. 29C), and the like. The storage tubes 884 are also referred to herein as vials 884 or cryovials 884 when used for storage and transport in cryogenic conditions. Examples of flasks include spinner flasks (not shown), multilayer flasks 350 (Millipore™ Millicell HY 3-layer cell culture flask T-600) shown in Fig. 20A, and the like. Examples of cell processing bottles include roller bottles (not shown) and the like. It should be understood that the above examples are not intended to be limiting and the term cell processing containers 314 as used herein could encompass any type of containers which are known to be

used for storing, treating, expanding and transporting batches. The ACPS 100 is also configured for the robotic handling of various types of reagent containers such as the reagent bottle 836 shown in Fig. 16.

[00153] As can be seen best in Figs. 4 and 5, the storage area 300 includes a left storage module 310, a central storage module 320, and a right storage module 330.

[00154] In the illustrated example stacking arrangement, the left storage module 310 holds stacks of carriers 312 for containers used for processing cells as can be seen best in Fig. 6. The left storage module includes a 9x3 array of carriers 312, each carrier 312 being capable of holding eight cell processing trays 344, 344' (Fig. 29B). The stackable carriers 312 allow the multiple cell processing trays 344 to be moved and stored together. The ACPS 100 also provides for cell processing containers 314 and reagent containers such as reagent bottles 836to be stored at, or subject to, temperatures below -100°C to +100°C, and be kept in the dark if needed.

[00155] In the illustrated example stacking arrangement, the right storage module 330 is configured to hold labware for cell processing as can be seen best in Fig. 6. The right storage module 330 includes five shelves 332 for storing labware with each shelf having five discrete positions 334 or trays 334 for holding labware. The labware stored in the shelves 332 of the right storage module 330 can be accessed (removed from shelf or placed thereon) in a random access manner. The vertical spacing between the consecutive shelves 332 of the right storage module 330 is not uniform in order to provide storage for labware of different heights.

WE CLAIM

1. A system for automated processing of batches, the batches being derived from biological samples, comprising:

a closed and sterile enclosure;

at least one station within the enclosure for holding a plurality of reagent containers;

at least one robotic reagent dispenser within the enclosure;

a quality control module within the enclosure for analyzing at least one characteristic of a batch;

a harvesting module within the enclosure;

a robotic module within the enclosure; and

a control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling said automatic processing of said batches, said automatic processing being executable without handling by a human operator.

2. The system of claim 1, further comprising a particle counter communicatively coupled to the CU for measuring a particle count inside the enclosure.

3. The system of claim 1 or 2, further comprising:

an air flow system for controlling an air pressure inside the enclosure to be greater than an air pressure outside the enclosure.

4. The system of any one of claims 1 to 3, wherein the enclosure is at least a class 10 environment.

5. The system of any one of claims 1 to 4, wherein:

the enclosure is defined at least in part by a plurality of walls including a top wall, a first side wall and a second side wall extending opposite the first side wall; the enclosure has an air inlet port defined in one or more of the plurality of walls of the enclosure;

the enclosure has an air outlet port defined in one or more of the plurality of walls of the enclosure,

the air flow system being configured to direct air flow into the enclosure via the air inlet port and air flow out of the enclosure via the air outlet port,

an air flow within the enclosure being laminar.

6. The system of any one of claims 1 to 5, wherein the system complies with good manufacturing practice (GMP) guidelines.

7. The system of any one of claims 1 to 6, further comprising a tracking module communicatively coupled to the CU for electronically tracking the batch after its introduction to the enclosure.

8. The system of any one of claims 1 to 7, wherein:

the CU is configured to control automatic processing of batches based on a signal received from the quality control module.

9. The system of any one of claims 1 to 8, wherein the batch comprises a plurality of batches, the system being configured to automatically process the plurality of batches without cross-contamination between batches.

10. The system of claim 9, wherein each of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module is configured to operate on any one of a first batch and a second batch such that:

when the first batch is being operated on by one of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module, the second batch is being operated on by an other of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module.

11. The system of claim 9, further configured to:

receive a second batch into the enclosure before a first batch is transported out of the enclosure; and

sequentially process each of the first batch and the second batch without cross contamination from the other of the first batch and the second batch.

12. The system of claim 11, further comprising a particle sensor communicatively coupled to the CU for measuring a particle count inside the enclosure, wherein:

the CU is configured to allow receiving of the second batch into the enclosure responsive to the particle count satisfying a predetermined criterion.

13. The system of claim 11 or 12, further comprising:

an automatic sterilizing module communicatively coupled to the CU for automatically sterilizing the enclosure before inserting the second batch into the enclosure.

14. The system of any one of claims 1 to 13, further comprising:

a waste receptacle selectively fluidly connected to the enclosure at a location remote from a center of the closed enclosure.

14A. The system of any one of claims 1 to 13, further comprising:

a waste receptacle selectively fluidly connected to the enclosure in a configuration that prevents particles in the waste receptacle from entering the enclosure.

15. The system of claim 14 or 14 A, wherein an air pressure in the waste receptacle is controlled to be lower than an air pressure inside the enclosure.

16. The system of claim 14, 14A or 15, wherein the enclosure is connected to the waste receptacle by a conduit configured to prevent backsplashing of waste material deposited into the conduit for disposal into the waste receptacle.

The system of any one of claims 1 to 16, further comprising:

an incubator selectively fluidly connected to the enclosure; and

a gate between the enclosure and the incubator being configured to be open for automatic transferring of the batch from the enclosure to the incubator and for automatic transferring of the batch from the incubator to the enclosure,

the enclosure being fluidly connected to the incubator when the gate is open and the enclosure being sealed from the incubator when the gate is closed.

18. The system of any one of claims 1 to 17, further comprising:

a centrifuge selectively fluidly connected to the enclosure; and

a gate between the enclosure and the centrifuge being configured to be open for automatic transferring of the batch from the enclosure to the centrifuge and for automatic transferring of the batch from the centrifuge to the enclosure,

the enclosure being fluidly connected to the centrifuge when the gate is open and the enclosure being sealed from the centrifuge when the gate is closed.

19. The system of any one of claims 1 to 18, wherein the robotic module comprises a plurality of robotic modules.

20. The system of any one of claims 1 to 19, wherein the robotic module is configured for at least one of:

transporting a container;

decapping or otherwise opening the container;

pipetting a reagent or liquid from the container; and

aspirating liquid from the container.

20A. The system of claim 20, wherein the liquid is cell culture media.

21. The system of any one of claims 1 to 20A, wherein one of the plurality of reagent containers is placed outside of the enclosure and connected to one of the at least one reagent dispensers by a fill line.

22. The system of any one of claims 1 to 21, further comprising a tilt module configured to tilt a container.

23. The system of claim 22, wherein the tilt module is a magnetic tilt module for separating magnetically tagged contents in the container from magnetically untagged contents of the container while tilting the container.

24. The system of any one of claims 1 to 23, wherein the quality control module is one of:

a flow cytometer;

a plate reader;

a microscope; and

a PCR machine.

25. The system of any one of claims 1 to 24, wherein the quality control module comprises a plurality of quality control modules.

26. The system of any one of claims 1 to 25, wherein the harvesting module comprises at least one of:

a container for holding a solution;

a freezer or a cryofreezer; and

a packaging module configured to place the batch in a transport container.

27. The system of any one of claims 1 to 26, further comprising an isolator,

the enclosure being selectively fluidly connected to the isolator, and objects from outside the system being received into the enclosure via the isolator, and/or

objects from inside the enclosure being passed out of the system via the isolator.

28. The system of claim 27, wherein:

an air pressure inside the enclosure is greater than an air pressure in the isolator; and

an air pressure inside the isolator is greater than an air pressure adjacent to the isolator in a direction other than the enclosure, or an ambient air pressure outside the system.

29. The system of claim 27 or 28, further comprising a biological safety cabinet (BSC),

the isolator being selectively fluidly connected to the BSC, and

objects from outside the system being received into the isolator via the BSC, objects from inside the enclosure being passed out of the system by passing from the enclosure to the isolator and from the isolator to the BSC via the isolator.

30. The system of any one of claims 1 to 29, further comprising:

one or more of a refrigerator and a freezer selectively fluidly connected to the enclosure; and

a gate between the enclosure and the one or more refrigerator or freezer being configured to be open for automatic transferring of the batch from the enclosure to the freezer and for automatic transferring of the batch from the one or more refrigerator or freezer to the enclosure,

the enclosure being fluidly connected to the one or more refrigerator or freezer when the gate is open and the enclosure being sealed from the one or more refrigerator or freezer when the gate is closed.

31. The system of any one of claims 1 to 30, wherein:

the enclosure is a first enclosure;

the plurality of reagent containers is a plurality of first reagent containers; at least one reagent dispenser is an at least one first reagent dispenser;

the quality control module is a first quality control module for analyzing at least one characteristic of a first batch;

the harvesting module is a first harvesting module; and

the robotic module is a first robotic module;

the system further comprising:

an incubator disposed outside the first enclosure, the first enclosure being selectively fluidly connected to the incubator;

a second closed and sterile enclosure selectively fluidly connected to the first enclosure or to the incubator;

a plurality of second reagent containers;

at least one second reagent dispenser;

a second quality control module for analyzing at least one characteristic of a second batch;

a second harvesting module; and

a second robotic module,

the CU being communicatively coupled to the at least one first and second reagent dispenser, the first and second quality control module, the first and second harvesting module and the first and second robotic module for controlling automatic processing of the first and second batches without handling by a human operator.

32. The system of any one of claims 1 to 30, wherein the system is selectively fluidly connected to a second system for automated processing of batches derived from biological samples, the second system comprising:

a second closed and sterile enclosure;

a second plurality of reagent containers;

a second at least one reagent dispenser;

a second quality control module for analyzing at least one characteristic of a batch;

a second harvesting module;

a second robotic module; and

a second control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling said automatic processing of said batches, said automatic processing being executable without handling by a human operator;

the CU and the second CU optionally being communicatively coupled to each other.

33. The system of claim 32, wherein the system and the second system are selectively fluidly connected via a second freezer or a second incubator, the second freezer or the second incubator being disposed outside both the enclosure and the second enclosure and being selectively fluidly connected to each of the enclosure and the second enclosure.

34. A system for automated processing of a plurality of batches, the batches being derived from biological samples, comprising:

a closed and sterile enclosure;

a plurality of reagent containers within the enclosure;

at least one reagent dispenser within the enclosure;

a quality control module within the enclosure for analyzing at least one characteristic of a batch;

a harvesting module within the enclosure;

a robotic module within the enclosure; and

a control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling said automatic processing of said batches,

the system being configured to automatically process the plurality of batches without cross-contamination between batches.

35. The system of claim 34, wherein the system is configured to automatically process the plurality of batches at the same time using sequential processing.

36. The system of claim 34 or 35, wherein the system is configured to automatically process the plurality of batches in compliance with good manufacturing practice (GMP) guidelines.

37. The system of any one of claims 34 to 36, wherein said automatic processing is executable without handling by a human operator.

38. The system of any one of claims 34 to 37, further comprising a particle counter communicatively coupled to the CU for measuring a particle count inside the enclosure.

39. The system of any one of claims 34 to 38, further comprising:

an air flow system for controlling an air pressure inside the enclosure to be greater than an air pressure outside the enclosure.

40. The system of any one of claims 34 to 39, wherein the enclosure is at least a class 10 environment.

41. The system of any one of claims 34 to 40, wherein:

the enclosure is defined at least in part by a top wall, a first side wall and a second side wall extending opposite the first side wall

the enclosure has an air inlet port defined in a top wall;

the enclosure has an air outlet port disposed in each of the first and second side walls,

the air flow system being configured to direct air flow into the enclosure via the air inlet port and air flow out of the enclosure via the air outlet port,

an air flow within the enclosure being laminar.

42. The system of any one of claims 34 to 41, wherein the system complies with good manufacturing practice (GMP) guidelines.

43. The system of any one of claims 34 to 42, further comprising a tracking module communicatively coupled to the CU for electronically tracking the batch after its introduction to the enclosure.

44. The system of any one of claims 34 to 43, wherein:

the CU is configured to control automatic processing of batches based on a signal received from the quality control module.

45. The system of any one of claims 34 to 44, wherein each of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module is configured to operate on any one of a first batch and a second batch such that:

when the first batch is being operated on by one of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module, the second batch is being operated on by an other of the at least one reagent dispenser, the quality control module, the harvesting module, and the robotic module.

46. The system of claim 45, further configured to:

receive a second batch into the enclosure before a first batch is transported out of the enclosure; and

sequentially process each of the first batch and the second batch without cross contamination from the other of the first batch and the second batch.

47. The system of claim 46, further comprising a particle sensor communicatively coupled to the CU for measuring a particle count inside the enclosure, wherein:

the CU is configured to allow receiving of the second batch into the enclosure responsive to the particle count satisfying a predetermined criterion.

48. The system of any one of claims 34 to 47, further comprising:

an automatic sterilizing module communicatively coupled to the CU for automatically sterilizing the enclosure before inserting the second batch into the enclosure.

49. The system of any one of claims 34 to 48, further comprising:

a waste receptacle selectively fluidly connected to the enclosure at a location remote from a center of the closed enclosure.

49A. The system of any one of claims 34 to 48, further comprising:

a waste receptacle selectively fluidly connected to the enclosure in a configuration that prevents particles in the waste receptacle from entering the enclosure.

50. The system of claim 49 or 49A, wherein an air pressure in the waste receptacle is controlled to be lower than an air pressure inside the enclosure.

51. The system of claim 49 or 49A or 50, wherein the enclosure is connected to the waste receptacle by a conduit configured to prevent backsplashing of waste material deposited into the conduit for disposal into the waste receptacle.

52. The system of any one of claims 34 to 51, further comprising:

an incubator selectively fluidly connected to the enclosure; and

a gate between the enclosure and the incubator being configured to be open for automatic transferring of the batch from the enclosure to the incubator and for automatic transferring of the batch from the incubator to the enclosure,

the enclosure being fluidly connected to the incubator when the gate is open and the enclosure being sealed from the incubator when the gate is closed.

53. The system of any one of claims 34 to 52, further comprising:

a centrifuge selectively fluidly connected to the enclosure; and

a gate between the enclosure and the centrifuge being configured to be open for automatic transferring of the batch from the enclosure to the centrifuge and for automatic transferring of the batch from the centrifuge to the enclosure,

the enclosure being fluidly connected to the centrifuge when the gate is open and the enclosure being sealed from the centrifuge when the gate is closed.

54. The system of any one of claims 34 to 53, wherein the robotic module comprises a plurality of robotic modules.

55. The system of any one of claims 34 to 54, wherein the robotic module is configured for at least one of:

transporting a container;

decapping or otherwise opening the container;

pipetting a reagent or liquid from the container; and

aspirating a liquid from the container.

55A. The system of claim 55, wherein the liquid is cell culture media.

56. The system of any one of claims 34 to 55A, wherein one of the plurality of reagent containers is placed outside of the enclosure and connected to one of the at least one reagent dispensers by a fill line.

57. The system of any one of claims 34 to 56, further comprising a tilt module configured to tilt a container.

58. The system of claim 57, wherein the tilt module is a magnetic tilt module for separating magnetically tagged contents in the container from magnetically untagged contents of the container while tilting the container.

59. The system of any one of claims 34 to 58, wherein the quality control module is one of:

a flow cytometer;

a plate reader;

a microscope; and

a PCR machine.

60. The system of any one of claims 34 to 59, wherein the quality control module comprises a plurality of quality control modules.

61. The system of any one of claims 34 to 60, wherein the harvesting module comprises at least one of:

a container for holding a solution;

a freezer or a cryofreezer; and

a packaging module configured to place the batch in a transport container.

62. The system of any one of claims 34 to 61, further comprising an isolator,

the enclosure being selectively fluidly connected to the isolator, and objects from outside the system being received into the enclosure via the isolator, and/or

objects from inside the enclosure being passed out of the system via the isolator.

63. The system of claim 62, wherein:

an air pressure inside the enclosure is greater than an air pressure in the isolator; and

an air pressure inside the isolator is greater than an air pressure adjacent to the isolator in a direction other than the enclosure, or an ambient air pressure outside the system.

64. The system of claim 62 or 63, further comprising a biological safety cabinet (BSC),

the isolator being selectively fluidly connected to the BSC, and

objects from outside the system being received into the isolator via the BSC, objects from inside the enclosure being passed out of the system by passing from the enclosure to the isolator and from the isolator to the BSC via the isolator.

65. The system of any one of claims 34 to 64, further comprising:

one or more of a refrigerator and a freezer selectively fluidly connected to the enclosure; and

a gate between the enclosure and the one or more refrigerator or freezer being configured to be open for automatic transferring of the batch from the enclosure to the freezer and for automatic transferring of the batch from the one or more refrigerator or freezer to the enclosure,

the enclosure being fluidly connected to the one or more refrigerator or freezer when the gate is open and the enclosure being sealed from the one or more refrigerator or freezer when the gate is closed.

66. The system of any one of claims 34 to 65, wherein:

the enclosure is a first enclosure;

the plurality of reagent containers is a plurality of first reagent containers; at least one reagent dispenser is an at least one first reagent dispenser;

the quality control module is a first quality control module for analyzing at least one characteristic of a first batch;

the harvesting module is a first harvesting module; and

the robotic module is a first robotic module;

the system further comprising:

an incubator disposed outside the first enclosure, the first enclosure being selectively fluidly connected to the incubator;

a second closed and sterile enclosure selectively fluidly connected to the first enclosure or to the incubator;

a plurality of second reagent containers;

at least one second reagent dispenser;

a second quality control module for analyzing at least one characteristic of a second batch;

a second harvesting module; and

a second robotic module,

the CU being communicatively coupled to the at least one first and second reagent dispenser, the first and second quality control module, the first and second harvesting module and the first and second robotic module for controlling automatic processing of the first and second batches without handling by a human operator.

67. The system of any one of claims 34 to 66, wherein the system is selectively fluidly connected to a second system for automated processing of batches derived from biological samples, the second system comprising:

a second closed and sterile enclosure;

a second plurality of reagent containers;

a second at least one reagent dispenser;

a second quality control module for analyzing at least one characteristic of a batch;

a second harvesting module;

a second robotic module; and

a second control unit (CU) communicatively coupled to the at least one reagent dispenser, the quality control module, the harvesting module and the robotic module for controlling said automatic processing of said batches, said automatic processing being executable without handling by a human operator;

the CU and the second CU optionally being communicatively coupled to each other.

68. The system of claim 67, wherein the system and the second system are selectively fluidly connected via a second freezer or a second incubator, the second freezer or the second incubator being disposed outside both the enclosure and the second enclosure and being selectively fluidly connected to each of the enclosure and the second enclosure.

69. The system of any one of claims 1 to 68 further comprising a robotic arm configured to receive a sample into the enclosure.

70. The system of any one of claims 1 to 69, further comprising a quality assurance module.

71. An automated method for processing a batch in a closed and sterile enclosure, the batch being derived from a biological sample inserted into the enclosure, the automated method comprising:

automatically processing the batch with one or more reagents;

automatically analyzing at least one characteristic of the batch; and

after automatically processing the batch, automatically harvesting the batch for reception outside the enclosure;

the automated method being executable without any handling by a human operator.

72. The method of claim 71 further comprising measuring a particle count inside the enclosure.

73. The method of claim 71 or 72, further comprising:

controlling an air pressure inside the enclosure to be greater than an air pressure outside the enclosure.

74. The method of any one of claims 71 to 73, further comprising electronically tracking the batch.

75. The method of any one of claims 71 to 74, further comprising:

determining a process rate based on the automatically analyzed at least one characteristic.

76. The method of any one of claims 71 to 75, wherein the batch comprises a plurality of batches, and the method comprises automatically processing each of the plurality of batches without cross -contamination between batches.

77. The method of claim 76, wherein the plurality of batches comprises:

a first batch including cells of a first type when inserted into the enclosure; and a second batch including cells of a second type when inserted into the enclosure.

78. The method of claim 77, wherein the first type is different from the second type.

79. The method of claim 77, wherein the first type is the same as the second type.

80. The method of any one of claims 77 to 79, wherein:

the automatically processing of the first batch is for transforming cells of the first type into cells of a third type; and

the automatically processing of the second batch is for transforming cells of the second type into cells of a fourth type.

81. The method of claim 80, wherein the third type is different from the fourth type.

82. The method of claim 80, wherein the third type is the same as the fourth type.

83. The method of any one of claims 71 to 76, wherein the batch is a first batch, and the method further comprises:

inserting a second batch into the enclosure;

automatically processing the second batch within the enclosure without interrupting the automatic processing of the first batch and without cross contamination from the first batch;

automatically analyzing at least one characteristic of the second batch; and automatically harvesting the second batch after the automatically processing.

84. The method of claim 83, wherein:

the second batch is inserted into the enclosure before the first batch is harvested.

85. The method of claim 83 or 84, further comprising measuring a particle count inside the enclosure, wherein:

the second batch is inserted into the enclosure responsive to the particle count satisfying a predetermined criterion.

86. The method of any one of claims 83 to 85, further comprising measuring a particle count inside the enclosure wherein:

the first batch and the second batch are automatically processed based on the particle count satisfying a predetermined criterion

87. The method of any one of claims 83 to 86, further comprising:

automatically sterilizing the enclosure before inserting the second batch into the enclosure.

88. The method of any one of claims 71 to 87, further comprising:

automatically disposing waste material in a waste receptacle selectively fluidly connected to the enclosure at a location remote from a majority of processing stations in the enclosure.

89. The method of claim 88, further comprising:

controlling an air pressure inside the waste receptacle to be lower than an air pressure inside the closed enclosure.

90. The method of any one of claims 71 to 89, further comprising:

automatically transferring the batch from the enclosure to a closed and sterile incubator, the enclosure being selectively fluidly connected to the incubator;

incubating the at least one batch;

automatically transferring the batch from the incubator to the closed enclosure; and

controlling a gate between the enclosure and the incubator to be open during the automatic transferring of the batch from the enclosure to the incubator and the automatic transferring of the batch from the incubator to the enclosure, the enclosure being fluidly connected to the incubator when the gate is open and the closed enclosure being sealed from the incubator when the gate is closed.

91. The method of any one of claims 71 to 90, further comprising:

automatically transferring the batch from the enclosure to a closed and sterile centrifuge, the enclosure being selectively fluidly connected to the centrifuge;

centrifuging the at least one batch;

automatically transferring the at least one batch from the centrifuge to the closed enclosure; and

controlling a gate between the enclosure and the centrifuge to be open during the automatic transferring of the batch from the enclosure to the centrifuge and the automatic transferring of the batch from the centrifuge to the enclosure, the closed enclosure being fluidly connected to the centrifuge when the gate is open and the closed enclosure being sealed from the centrifuge when the gate is closed.

92. The method of any one of claims 71 to 91, wherein the one or more reagents comprises a reprogramming agent.

93. The method of claim 92, wherein the reprogramming agent comprises a small molecule, a chemical compound, a peptide, or a polynucleotide encoding a peptide.

94. The method of claim 92 or 93, wherein the reprogramming agent is a gene regulator or acts to increase expression of a gene regulator in cells in the batch.

95. The method of any one of claims 71 to 94, further comprising:

automatically transferring the batch from a first processing station to a second processing station.

96. The method of any one of claims 71 to 95, wherein automatically processing the batch with one or more reagents comprises:

automatically performing a passaging protocol for placing the batch from a first cell processing container to a plurality of second cell processing containers.

97. The method of any one of claims 71 to 96, wherein automatically harvesting comprises:

automatically placing the batch into a transport container.

98. The method of any one of claims 71 to 97, wherein automatically harvesting comprises:

freezing or cryofreezing the batch.

99. The method of claim 98, wherein cryofreezing comprises nucleated cryofreezing.

100. The method of any one of claims 71 to 99, wherein the automatically analyzing at least one characteristic of the batch comprises analyzing at least one of: a cell confluency;

a cell number;

a cell viability;

a cell density;

a cell diameter;

a marker expression;

a morphological characteristic;

a cell differentiation characteristic;

a cell potency;

a cell purity;

a cell sterility; and

a cell identity;

101. The method of any one of claims 71 to 100, wherein at least one characteristic of the batch is automatically analyzed before completion of said automatically processing the batch with said one or more reagents.

102. The method of any one of claims 71 to 101, wherein at least one characteristic of the batch is automatically analyzed after completion of said processing the batch with said one or more reagents.

103. The method of any one of claims 71 to 102, wherein at least one characteristic of the batch is automatically analyzed before harvesting the batch.

104. The method of any one of claims 71 to 103, wherein the method is executed in compliance with good manufacturing practice (GMP) guidelines.

105. The method of any one of claims 71 to 104, further comprising controlling an environment in the enclosure to maintain the enclosure as at least a class 10 environment.

106. The method of any one of claims 71 to 105, further comprising:

receiving objects into the enclosure via an isolator selectively fluidly connected to the enclosure, the isolator being sterile and closed.

107. The method of claim 106, further comprising:

sterilizing objects in the isolator before passing objects from the isolator into the enclosure.

108. The method of claim 106 or 107, further comprising:

passing objects from the enclosure to an outside of the enclosure via the isolator.

109. The method of any one of claims 106 to 108, further comprising:

receiving objects into a biological safety cabinet (BSC) before passing the objects from the BSC into the isolator and via the isolator into the enclosure,

the BSC being selectively fluidly connected to the isolator, the BSC being sterile and closed.

110. The method of claim 109, further comprising:

passing objects from the isolator or a second isolator to an outside of the enclosure via the biological safety cabinet (BSC or a second biological safety cabinet.

111. An automated method for processing a batch in a closed and sterile enclosure, the batch being derived from a biological sample inserted into the enclosure, the automated method comprising:

automatically processing the batch with one or more reagents;

automatically analyzing at least one characteristic of the batch; and after automatically processing the batch, automatically harvesting the batch for reception outside the enclosure;

wherein the automated method is capable of processing a plurality of batches without cross-contamination between batches.

112. The method of claim 111, wherein the plurality of batches are processed at the same time using sequential processing.

113. The method of claim 111 or 112, wherein the plurality of batches are processed in compliance with good manufacturing practice (GMP) guidelines.

114. The method of any one of claims 111 to 113, wherein the automated method is executable without any handling by a human operator.

115. The method of any one of claims 111 to 114, further comprising measuring a particle count inside the enclosure.

116. The method of any one of claims 111 to 115, further comprising:

controlling an air pressure inside the enclosure to be greater than an air pressure outside the enclosure.

117. The method of any one of claims 111 to 116, further comprising electronically tracking the batch.

118. The method of any one of claims 111 to 117, further comprising:

determining a process rate based on the automatically analyzed at least one characteristic.

119. The method of any one of claims 111 to 118, wherein the plurality of batches comprises:

a first batch including cells of a first type when inserted into the enclosure; and a second batch including cells of a second type when inserted into the enclosure.

120. The method of claim 119, wherein the first type is different from the second type.

121. The method of claim 119, wherein the first type is the same as the second type.

122. The method of any one of claims 119 to 121, wherein:

the automatically processing of the first batch is for transforming cells of the first type into cells of a third type; and

the automatically processing of the second batch is for transforming cells of the second type into cells of a fourth type.

123. The method of claim 122, wherein the third type is different from the fourth type.

124. The method of claim 122, wherein the third type is the same as the fourth type.

125. The method of any one of claims 111 to 124, wherein the batch is a first batch, and the method further comprises:

inserting a second batch into the enclosure;

automatically processing the second batch within the enclosure without interrupting the automatic processing of the first batch and without cross contamination from the first batch;

automatically analyzing at least one characteristic of the second batch; and automatically harvesting the second batch after the automatically processing.

126. The method of claim 125, wherein:

the second batch is inserted into the enclosure before the first batch is harvested.

127. The method of claim 125 or 126, further comprising measuring a particle count inside the enclosure, wherein:

the second batch is inserted into the enclosure responsive to the particle count satisfying a predetermined criterion.

128. The method of any one of claims 111 to 127, further comprising measuring a particle count inside the enclosure wherein:

the first batch and the second batch are automatically processed based on the particle count satisfying a predetermined criterion

129. The method of any one of claims 111 to 128, further comprising:

automatically sterilizing the enclosure before inserting the second batch into the enclosure.

130. The method of any one of claims 111 to 129, further comprising:

automatically disposing waste material in a waste receptacle selectively fluidly connected to the enclosure at a location remote from a majority of processing stations in the enclosure.

131. The method of claim 130, further comprising:

controlling an air pressure inside the waste receptacle to be lower than an air pressure inside the closed enclosure.

132. The method of any one of claims 111 to 131, further comprising:

automatically transferring the batch from the enclosure to a closed and sterile incubator, the enclosure being selectively fluidly connected to the incubator;

incubating the at least one batch;

automatically transferring the batch from the incubator to the closed enclosure; an

controlling a gate between the enclosure and the incubator to be open during the automatic transferring of the batch from the enclosure to the incubator and the automatic transferring of the batch from the incubator to the enclosure, the enclosure being fluidly connected to the incubator when the gate is open and the closed enclosure being sealed from the incubator when the gate is closed.

133. The method of any one of claims 111 to 132, further comprising:

automatically transferring the batch from the enclosure to a closed and sterile centrifuge, the enclosure being selectively fluidly connected to the centrifuge;

centrifuging the at least one batch;

automatically transferring the at least one batch from the centrifuge to the closed enclosure; and

controlling a gate between the enclosure and the centrifuge to be open during the automatic transferring of the batch from the enclosure to the centrifuge and the automatic transferring of the batch from the centrifuge to the enclosure, the closed enclosure being fluidly connected to the centrifuge when the gate is open and the closed enclosure being sealed from the centrifuge when the gate is closed.

134. The method of any one of claims 111 to 133, wherein the one or more reagents comprises a reprogramming agent.

135. The method of claim 134, wherein the reprogramming agent comprises a small molecule, a chemical compound, a peptide, or a polynucleotide encoding a peptide.

136. The method of claim 134 or 135, wherein the reprogramming agent is a gene regulator or acts to increase expression of a gene regulator in cells in the batch.

137. The method of any one of claims 111 to 136, further comprising:

automatically transferring the batch from a first processing station to a second processing station.

138. The method of any one of claims 111 to 137, wherein automatically processing the batch with one or more reagents comprises:

automatically performing a passaging protocol for placing the batch from a first cell processing container to a plurality of second cell processing containers.

139. The method of any one of claims 111 to 138, wherein automatically harvesting comprises:

automatically placing the batch into a transport container.

140. The method of any one of claims 111 to 139, wherein automatically harvesting comprises:

cryofreezing the batch.

141. The method of claim 140, wherein cryofreezing comprises nucleated cryofreezing.

142. The method of any one of claims 111 to 141, wherein the automatically analyzing at least one characteristic of the batch comprises analyzing at least one of: a cell confluency;

a cell number;

a cell viability;

a cell density;

a cell diameter;

a marker expression;

a morphological characteristic;

a cell differentiation characteristic;

a cell potency;

a cell purity;

a cell sterility; and

a cell identity;

143. The method of any one of claims 111 to 142, wherein at least one characteristic of the batch is automatically analyzed before completion of said automatically processing the batch with said one or more reagents.

144. The method of any one of claims 111 to 143, wherein at least one characteristic of the batch is automatically analyzed after completion of said processing the batch with said one or more reagents.

145. The method of any one of claims 111 to 144, wherein at least one characteristic of the batch is automatically analyzed before harvesting the batch.

146. The method of any one of claims 111 to 145, wherein the method is executed in compliance with good manufacturing practice (GMP) guidelines.

147. The method of any one of claims 111 to 146, further comprising controlling an environment in the enclosure to maintain the enclosure as at least a class 10 environment.

148. The method of any one of claims 111 to 147, further comprising:

receiving objects into the enclosure via an isolator selectively fluidly connected to the enclosure, the isolator being sterile and closed.

149. The method of claim 148, further comprising:

sterilizing objects in the isolator before passing objects from the isolator into the enclosure.

150. The method of claim 148 or 149, further comprising:

passing objects from the enclosure to an outside of the enclosure via the isolator.

151. The method of any one of claims 148 to 150, further comprising:

receiving objects into a biological safety cabinet (BSC) before passing the objects from the BSC into the isolator and via the isolator into the enclosure,

the BSC being selectively fluidly connected to the isolator, the BSC being sterile and closed.

152. The method of claim 151, further comprising:

passing objects from the isolator or a second isolator to an outside of the enclosure via the biological safety cabinet (BSC) or a second biological safety cabinet.

153. The method of any one of claims 71 to 152 further comprising:

introducing the sample into the enclosure;

automatically preparing the batch from the sample after introduction of the sample and before processing of the batch.

154. The method of claim 153, wherein the sample is introduced into the enclosure robotically.

155. The method of any one of claims 71 to 154, further comprising:

automatically packaging the batch for storage or transport.

156. The method of any one of claims 71 to 155, further comprising:

automatically sterilizing the enclosure.

157. The method of any one of claims 71 to 156, further comprising:

automatically conducting quality assurance analysis.

158. The method of claim 157, wherein the quality assurance analysis comprises reviewing the steps executed for the batch and determining if pre-determined criteria are met.

159. The method of claim 157 or 158, wherein the quality assurance analysis comprises testing for sterility, endotoxin, or mycoplasma.

160. The method of any one of claims 157 to 159, wherein the pre-determined criteria are GMP regulations.

161. The method of any one of claims 157 to 160, further comprising automatically producing a quality assurance report.

162. A method for automated sterilization of an enclosure for processing of batches comprising:

injecting sterilant into the enclosure via a sterilant inlet;

circulating within the enclosure sterilant received via the sterilant inlet;

removing, from the enclosure, the sterilant circulating within the enclosure; closing at least one port of the enclosure other than the sterilant inlet and sterilant outlet before injecting sterilant into the enclosure and keeping the at least one port closed while injecting sterilant into the enclosure; and

removing from the enclosure, or sealing within a sub-enclosure of the enclosure, any container containing a portion of the batches before receiving sterilant into the enclosure.

163. The method of claim 162, wherein the at least one port is a refrigerator or freezer access port connecting a refrigerator or freezer disposed outside the enclosure to the enclosure and further comprising:

closing an insulation door of the refrigerator or freezer, the insulation door providing an additional layer of insulation between the enclosure and the refrigerator or freezer.

164. A robotic aspirator comprising:

a robotic arm configured to move in at least one direction;

a body connected to the robotic arm;

an aspiration member comprising a fluid flow channel connected to the body, the aspiration member being configured for connection to a pump means;

the body being configured to hold a disposable tip for providing fluid connection between the disposable tip and the fluid flow channel of the aspiration member;

fluid being aspirated through the disposable tip and the fluid flow channel when the disposable tip is fluidly connected to the fluid flow channel and the aspiration member is connected to the pump means.

165. The robotic aspirator of claim 164, further comprising a plurality of prongs connected to the body, the prongs being moveable between a tip holding position and a retracted position, the prongs being configured in the tip holding position to hold the disposable tip for providing fluid connection between the disposable tip and the fluid flow channel of the aspiration member.

166. The robotic aspirator of claim 165, the plurality of prongs being further configured for gripping objects other than the disposable tip.

167. A method of aspirating a sample using the robotic aspirator of any one of claims 164 to 166.

168. The system of any one of the preceding claims comprising the robotic aspirator of any one of claims 164 to 166.

169. The system of claim 168, comprising means for engaging and/or disengaging the disposable tip from the robotic aspirator without handling by a human operator.

170. The method of any one of the preceding claims comprising a step of aspirating a sample using the robotic aspirator of any one of claims 164 to 166.

171. A method of aspirating using a robotic arm having a fluid flow channel and a plurality of prongs configured to selectively hold a disposable tip in fluid connection with the fluid flow channel, the method comprising:

moving the prongs to retain the disposable tip in fluid connection with the fluid flow channel, the prongs being selectively moveable and optionally further configured to grip at least one object other than the disposable tip; and

evacuating the fluid flow channel to aspirate liquid through the disposable tip and the fluid flow channel.

172. The method of claim 171, further comprising:

after aspirating liquid, disengaging the prongs from the disposable tip; and

stopping evacuation of the fluid flow channel to disengage the disposable tip from the fluid flow channel.

173. The method of claim 172, wherein the disposable tip disengages from the fluid flow channel without handling by a human operator.

174. The method of any one of the preceding claims further comprising the method of any one of claims 171 to 173.

175. The system of any one of the preceding claims, comprising an automated liquid fill station housed inside the enclosure and configured for direct filling of liquid from a supply container disposed inside or outside the enclosure into a cell processing container disposed inside the enclosure, optionally wherein the liquid is media, optionally wherein the liquid fill station is configured to dispense more than 5 mL of liquid at a time.

176. The method of any one of the preceding claims, comprising a step of direct filling a liquid from a supply container disposed inside or outside the enclosure into a cell processing container disposed inside the enclosure, optionally wherein the liquid is media, optionally wherein more than 5 mL of liquid is dispensed at a time.

178. The system of any one of the preceding claims, comprising robotic means for tilting a cell processing container, for delivery of cells or media thereto and/or removal of cells or media therefrom.

179. The system of claim 178, wherein the cell processing container is exposed to a magnet during tilting, for magnetic sorting, separation or treatment of cells.

180. The system of any one of the preceding claims, comprising robotic means for capping and decapping screwcap lids of containers.

181. The system of claim 180, wherein the containers are sized to hold volumes of greater than 10 mL or greater than 2 mL.

182. The system of any one of the preceding claims, comprising automated means for purifying cell mixtures or obtaining a cell pellet.

183. The system of claim 182, wherein said means comprises a centrifuge.

184. The system of any one of the preceding claims, comprising means for incubating cells.

185. The system of any one of the preceding claims, comprising automated means for determining cell number or cell confluency in a sample.

186. The system of any one of the preceding claims, comprising a vertical waste chute for discarding solid waste configured so that the waste does not hit the edges of the chute, and placed under sufficient negative pressure such that particles from the chute do not enter the enclosure.

187. The system of any one of the preceding claims, comprising sterilization means for sterilizing the enclosure.

188. The system of claim 187, wherein the system is configured to allow sterilization of the enclosure without removing the batches from the system.

189. The system of any one of the preceding claims, comprising robotic means for freezing of batches and/or means for manipulation of samples or reagents at subzero temperatures.

190. The system of any one of the preceding claims, the system comprising an enclosure and one or more of the following automated components:

(1) a robotic aspirator housed within the enclosure and configured for use with disposable tips, as described in any one of claims 164 to 166;

(2) a decapper housed within the enclosure and configured to decap a screwcap lid of a container, optionally wherein the container is of 10 mL or greater volume or of 2 mL or greater volume;

(3) a centrifuge, cell sorter, or magnet;

(4) an incubator;

(5) a module for determining confluency of cells and/or cell number;

(6) a liquid fill module housed inside the enclosure and configured for direct filling of liquid from a supply container disposed inside or outside the enclosure into a cell processing container disposed inside the enclosure, optionally wherein the liquid is media, optionally wherein the liquid fill station is configured to dispense more than 5 mL of liquid at a time; and

(7) a tilt module configured for tilting a cell processing container, for removal or collection of cells or media therefrom, optionally wherein the tilt module is magnetic.

191. The system of claim 190, wherein the system comprises two or more, three or more, four or more, five or more, more than 5 or all of the automated components (1) to (7).

192. The system of any one of the preceding claims, wherein the system further comprises a module for quality assurance.

193. A method for automated processing of cells, the method comprising the steps of:

providing a biological sample to a closed and sterile enclosure, the closed and sterile enclosure housing:

robotic means for receiving the biological sample;

robotic means for preparing cells for processing from the biological sample;

robotic means for processing the cells;

robotic means for analyzing at least one characteristic of a batch; and robotic means for harvesting the cells after processing; providing a control unit communicatively coupled to the enclosure for controlling each of said robotic means, and

the control unit executing a desired processing program in the enclosure, said automated processing being executed without handling by a human operator.

194. The method of claim 193, wherein the enclosure further houses robotic means for packaging, and optionally labelling the cells for storage and/or transport, after harvesting.

195. The method of claim 193 or 194, wherein the enclosure further houses means for quality assurance and/or automated pass/fail criteria for approving or not approving the release of a batch.

196. The method of any one of claims 193 to 195, wherein the enclosure, the robotic means and/or the control unit are configured to process a plurality of biological samples and cells prepared therefrom without cross -contamination.

197. The method of any one of claims 193 to 196, wherein said method is executed under conditions that meet GMP guidelines and regulations.

198. A biological sample prepared using the automated system or method of any one of the preceding claims.

199. The biological sample of claim 198, wherein the biological sample comprises a population of cells, a tissue matrix, an antibody, a protein, a biological compound, or a chemical compound.

200. The biological sample of claim 199, wherein the population of cells comprises multipotent, unipotent, somatic, or stem-like cells that have been obtained by reprogramming from another cell of a different type.

201. The biological sample of claim 199 or 200, wherein the population of cells comprises a population of in vitro human multipotent, unipotent, or somatic cells derived from the reprogramming of a somatic cell, a progenitor cell or a stem cell that exhibits at least a transient increase in intracellular levels of at least one reprogramming agent; wherein the multipotent, unipotent or somatic cells exhibit a stable, non-forced gene expression profile comprising (a) reduced expression of one or more markers specific to the somatic cell, progenitor cell, or stem cell from which the multipotent, unipotent, or somatic cell is derived; (b) maintained expression of one or more markers specific to the somatic cell, progenitor cell, or stem cell from which the multipotent, unipotent, or somatic cell is derived; and (c) expression of markers and characteristics of a reprogrammed multipotent, unipotent, or somatic cell; wherein the multipotent, unipotent or somatic cells are not differentiated from a pluripotent cell; wherein the multipotent, unipotent or somatic cells are not cancerous; and wherein the multipotent, unipotent or somatic cells do not exhibit uncontrolled growth, teratoma formation, and tumor formation.

202. The biological sample of any one of claims 199 to 201, wherein:

the population of cells comprises neural stem cells, neural stem-like cells, neural precursor cells, neural progenitor cells, neuroblasts, neurons, cardiac cells, hematopoietic cells, cells of ectoderm, mesoderm or endoderm lineage, pluripotent

cells, multipotent cells, unipotent cells, somatic cells, naturally occurring cells, non-naturally occurring cells, prokaryotic cells, and/or eukaryotic cells.

the population of cells comprises cells that can differentiate into a plurality of progenitor, precursor, or somatic cells;

the population comprises cells having the ability to proliferate for more than

30 population doublings, or cells capable of maintaining telomerase activity through to at least about 30 population doublings; and/or

the doubling time of the cell population is less than 7 days or less than 3 days.

203. A population of in vitro human multipotent, unipotent, or somatic cells derived from the reprogramming of a somatic cell, a progenitor cell or a stem cell of a different type using the automated system or method of any one of the preceding claims.

204. An automated process for reprogramming a cell of a first type to a desired cell of a different type that is multipotent or unipotent, the cell of a first type being a somatic cell, a stem cell, or a progenitor cell, the automated process executable by the system of any one of the preceding claims, the automated process comprising:

introducing into the cell of a first type using robotic means an agent capable of remodeling the chromatin and/or DNA of the cell, wherein the agent capable of remodeling the chromatin and/or DNA is a histone acetylator, an inhibitor of histone deacetylation, a DNA demethylator, and/or a chemical inhibitor of DNA methylation;

transiently increasing intracellular levels of at least one reprogramming agent in the cell of a first type using robotic means, wherein the at least one reprogramming agent increases directly or indirectly the endogenous expression of at least one multipotent or unipotent gene regulator to a level at which the gene regulator is capable of driving transformation of the cell of a first type into the multipotent or unipotent cell;

wherein the cell of a first type is maintained in culture conditions supporting the transformation of the cell of a first type to the multipotent or unipotent cell for a sufficient period of time to allow a stable expression of a plurality of secondary genes characteristic of the phenotypical and/or functional properties of the multipotent or unipotent cell, where one or more of the secondary genes is not characteristic of phenotypical and functional properties of an embryonic stem cell and wherein stable expression of the plurality of secondary genes occurs in the absence of the reprogramming agent, whereby at the end of said period of time the cell of a first type has been transformed into the multipotent or unipotent cell,

where the multipotent or unipotent cell expresses at least one marker characteristic of the cell of a first type.