Claims:What is claimed is:
1. A point of care tissue processing system for epidermal tissue harvesting cum dissociation, comprising:

an epidermal tissue harvesting and dissociation device is attached to a leg/hand of a person and tightened with at least one elastic adjustable band; at least one skin warming component is placed over the epidermal tissue harvesting and dissociation device and the least one skin warming component is configured to reach required temperature with at least one heating element inside the at least one skin warming component;

at least one control panel of a tissue processing instrument is configured to display temperature of the at least one skin warming component; a first port of the epidermal tissue harvesting and dissociation device is connected to the at least one vacuum pump of tissue processing instrument through at least one vacuum tube connector, whereby the at least one vacuum tube connector is configured to apply vacuum until a plurality of epidermal tissue blisters raised into a plurality of perforations in at least one tissue slicing assembly of the tissue harvesting and dissociation device, the plurality of epidermal tissue blisters is sliced by moving at least one tissue middle SS slicing plate horizontally with the help of at least one specific lever, the plurality of perforations is closed tightly with at least one silicone cap protrusions after the plurality of epidermal tissue blisters are sliced;

an enzyme (Trypsin: EDTA) solution is added to the epidermal tissue harvesting and dissociation device for a tissue dissociation through a second port after collection of the plurality of epidermal tissue blisters into the epidermal tissue harvesting and dissociation device and mix thoroughly, whereby the epidermal tissue harvesting and dissociation device is placed in at least one centrifuge rotor of the tissue processing instrument and tighten with a lid, the harvesting and dissociation device with the plurality of epidermal tissue blisters mixed by rotating the at least one centrifuge rotor in opposite directions under optimum temperature conditions in at least one rotor chamber until tissue is completely dissociated, the at least one 70 micron cell strainer of the epidermal tissue harvesting and dissociation device allows only dissociated cells to pass through to bottom of the epidermal tissue harvesting and dissociation device, the epidermal tissue harvesting and dissociation device is centrifuged to settle individual cells at bottom of the tissue processing instrument, an enzymatic solution is removed from the second port of the epidermal tissue harvesting and dissociation device; and

a cell washing buffer is added to the epidermal tissue harvesting and dissociation device through the second port and mixed by rotating the at least one centrifuge rotor forward and reverse directions, further, the individual cells are settled at the bottom of the tissue processing instrument by centrifugation, a single cell suspension is dissolved in the washing buffer and suspend the individual cells into the solution, the individual cell suspension is collected through the second port into an application syringe.

2. The point of care tissue processing system of claim 1, wherein the at least tissue slicing assembly and at least one dissociation assembly are configured to hold at least one of: at least one upper stainless steel slicing sheet; at least one middle stainless steel slicing sheet and at least one lower stainless steel slicing sheet.

3. The point of care tissue processing system of claim 2, wherein the at least one middle stainless steel slicing sheet is configured to move between the at least one upper stainless steel slicing sheet and the at least one lower stainless steel slicing sheet to slice the plurality of epidermal tissue blisters.

4. The point of care tissue processing system of claim 2, wherein the at least one middle stainless steel slicing sheet is extended outside and connected to the at least one specific lever to slice the plurality of epidermal tissue blisters.

5. The point of care tissue processing system of claim 1, wherein the tissue processing instrument comprising at least one vacuum pump and the at least one skin warming component that are used for the plurality of tissue blisters creation in the at least tissue slicing assembly.

6. The point of care tissue processing system of claim 5, wherein the at least vacuum pump’s pressure, temperature of the skin warming component are controlled by the at least one control panel in the tissue processing instrument.

7. The point of care tissue processing system of claim 1, wherein the tissue processing instrument comprising a plurality of temperature sensors, the at least one skin warming component, the at least one rotor chamber and vacuum sensors of the at least one vacuum pump connected to the at least one control panel in the tissue processing instrument.

8. The point of care tissue processing system of claim 2, wherein the at least one dissociation assembly comprising at least one oval shape container embedded with at least one cell strainer for enabling the individual cells to pass through pores after the tissue dissociation.

9. The point of care tissue processing system of claim 8, wherein the at least one 70 micron cell strainer comprising strainer teeth and the at least one dissociation assembly comprising the first port configured for vacuum creation and the second port configured to one of solution addition and solution removal from the at least one dissociation assembly.

10. The point of care tissue processing system of claim 1, wherein the tissue processing instrument comprising a plurality of Peltier modules configured to regulate the temperature of at least one rotor chamber.

11. The point of care tissue processing system of claim 1, wherein the at least one control panel is configured to control the at least one vacuum pump, the temperature of the at least one rotor chamber, the speed of the at least one centrifuge rotor, and the temperature of the at least one skin warming component.

12. The point of care tissue processing system of claim 1, wherein the at least one centrifuge rotor speed is fixed to a motor and placed inside the at least one rotor chamber.

13. The point of care tissue processing system of claim 12, wherein the motor is controlled to move the forward and reverse directions to mix tissue for dissociation along temperature maintenance in the at least one temperature controlled chamber.

14. The point of care tissue processing system of claim 1, wherein the dissociated cells are collected by pelleting by centrifugation in the tissue processing instrument.

15. A method for epidermal tissue harvesting cum dissociation, comprising:

attaching epidermal tissue harvesting and dissociation device onto a leg/hand of a user and tighten with at least one elastic adjustable band;

placing at least one skin warming component over the epidermal tissue harvesting and dissociation device and then adjusting the temperature inside the at least one skin warming component in at least one control panel of a tissue processing instrument;

connecting at least one first port of the epidermal tissue harvesting and dissociation device to the tissue processing instrument through a vacuum connector for vacuum creation;

forming a plurality of epidermal tissue blisters through a plurality of perforations and slicing the plurality of epidermal tissue blisters by moving tissue slicing plates outwards with the help of at least one specific lever, the plurality of epidermal tissue blisters are formed by creating vacuum in the tissue harvesting and dissociation device and applying temperature in the at least one skin warming component;

removing the epidermal tissue harvesting and dissociation device from the leg/hand and closing the plurality of perforations with a silicon cap;

adding an enzyme solution to the epidermal tissue harvesting and dissociation device and mixing the enzyme solution properly;

placing the epidermal tissue harvesting and dissociation device in a centrifuge rotor of the tissue processing instrument and tighten with at least one lid;

adjusting the temperature of at least one rotor chamber and selecting a mix option through the at least one control panel to mix tissues for tissue dissociation;

collecting and discarding the enzyme solution from the epidermal tissue harvesting and dissociation device after completion of dissociation;

adding a washing solution through a second port to at least one dissociation component, the washing solution configured to wash cells and centrifuging the at least one dissociation component to settle the cells at bottom of the epidermal tissue harvesting and dissociation device;

removing the washing solution and observing the cells are precipitated at the bottom of the epidermal tissue harvesting and dissociation device;

mixing the epidermal tissue harvesting and dissociation device in the tissue processing instrument to suspend the precipitated cells into the washing solution; and

collecting a cell suspension through a cell collection port into the syringe. , Description:TECHNICAL FIELD
[001] The disclosed subject matter relates generally to a device for harvesting a skin graft, and more particularly to appoint of care tissue processing system for epidermal tissue harvesting cum dissociation and method employed thereof.

BACKGROUND
[002] Skin grafting is a surgical procedure in which a section of skin is removed from one area of a patient's body (autograft) or removed from another human source (allograft), and transplanted to a recipient site of the patient’s body, such as a wound site or leukoderma. Autologous skin grafting procedures have become an almost integral part of advanced wound care (for example, treatment of ulcers, burns) Vitiligo/ leukoderma, etc.). Autologous epidermal tissue grafting is one of the superior treatment option for vitiligo /leukoderma, ulcers and burns patients. Harvesting of a skin graft may be accomplished by many different techniques, and the technique used may depend on the type of graft to be harvested. In the autologous tissue grafting procedure, intact epidermal tissue may be harvested in pieces format and grafted on the affected region. This method of tissue grafting techniques generally involve an equal amount of tissue being removed from a donor site and then graft onto required affected area.

[003] Newer methods of epidermal single “cell” suspension grafting rather “tissue” will cover larger area with less donor tissue harvest. The use of epidermal single cell suspension instead of intact epidermis is more effective and minimizes the donor area scars and engrafts easily. However this procedure require either dissociation of tissue or expensive and complex cultivation procedures using laboratory facilities to create larger portions of graft tissue from smaller donor samples.

[004] Epidermal single cell suspension preparation includes tissue harvesting from patients in the form of vacuum suction epidermal blisters and dissociate to make single cell suspension. Harvesting epidermis through suction blisters from humans is time taking process in regular formats and further making single cell suspension requires a large laboratory set up to avoid contamination during tissue dissociation. Combining all processes together requires the complete laboratory set up with biosafety cabinets and it takes at least 5-6 hours. This increases patient waiting periods and treatments costs.

[005] To improve the technique, developments happen to eases up the epidermis harvesting procedures using skin warming conditions. Warming the skin reduces the time for suction blisters formation thus reduces the time of harvesting. Following skin warming principle, certain devices can accomplish epidermis harvesting in tissue micro blister format and grafts directly to affected region. However, the existing technology do not address the tissue dissociation at point of care thus does not change the requirement of donor to recipient skin requirement ratio.

[006] Further, certain devices/ kits are available to make single cell suspension from tissues. However usage of these kits confined to specialized laboratories only. And also the process is difficult and costly for clinical approach as these kits are not designed for point of care clinical use.

[007] In the light of aforementioned discussion, there exists a need for a point of care tissue harvesting and processing device and instrumentation for executing the total procedure of epidermal cell grafting right from epidermal tissue harvesting to dissociation at clinical set up / point of care and method employed thereof.

BRIEF SUMMARY
[008] The following presents a simplified summary of the disclosure in order to provide a basic understanding to the reader. This summary is not an extensive overview of the disclosure and it does not identify key/critical elements of the invention or delineate the scope of the invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later.

[009] An objective of the present disclosure is directed towards a point of care tissue processing system that makes a single cell suspension from small quantity of tissue grafts for a large area of treatment, i.e. 1cm skin graft cells may be enough to cover 10cm of treatment area.

[0010] Another objective of the disclosure is directed towards a point of care system that can be handled by non-skilled personnel for preparation of single cell graft material from tissue harvest.

[0011] Another objective of the present disclosure is directed towards a point of care tissue processing system for epidermal tissue harvesting and dissociation to make single cell suspension which avoids requirement of large laboratory set up and cumbersome methodology thus reducing the cost of treatment.

[0012] Another objective of the present disclosure is directed towards a point of care tissue processing system that avoids contamination and saves time in preparation of epidermal cell suspension for treating the wounds, burns and vitiligo/leukoderma patients.

[0013] Yet another objective of the present disclosure is directed towards a point of care tissue processing system that can be used for other tissue (i.e. Dermis, cord tissue, pancreas, etc.) dissociation in “tissue in cells out” manner in research applications.
[0014] A point of care tissue processing system comprises of two components 1) a single use tissue harvesting and dissociation device and 2) Tissue processing instrument.

[0015] According to an to an exemplary embodiment of the present disclosure, the point of care tissue processing system comprising at least epidermal tissue harvesting and dissociation device is attached to a leg/hand of a person and tightened with at least one elastic adjustable band;

[0016] According to an exemplary disclosure, at least one skin warming component is placed over the at least one epidermal tissue harvesting and dissociation device and the least one skin warming component is configured to create required temperature of approx. 45°C with at least one heating element and temperature controlled by temperature control sensor placed inside the skin warming component that further connected to Tissue processing instrument control panel.

[0017] According to another exemplary embodiment of the present disclosure, the point of care tissue processing system comprising at least one control panel of at least one tissue processing instrument is configured to display and controls the temperature of the at least one skin warming component.

[0018] According to another exemplary embodiment of the present disclosure, a first port of the at least one epidermal tissue harvesting and dissociation device is connected to at least one vacuum pump of the tissue processing instrument through at least one vacuum tube connector, the at least one vacuum pump is configured to apply vacuum until a plurality of epidermal tissue blisters raised into a plurality of perforations in at least one tissue slicing assembly of the at least one tissue harvesting and dissociation device, the plurality of epidermal tissue blisters is sliced by moving at least one tissue slicing plate horizontally with the help of at least one specific lever, the plurality of perforations of the tissue slicing assembly is closed tightly with at least one silicone cap after the plurality of epidermal tissue blisters are sliced and collected into the tissue dissociation assembly.

[0019] According to another exemplary embodiment of the present disclosure, an enzyme solution is added to the at least one epidermal tissue harvesting and dissociation device for a tissue dissociation through a second port after collection of the plurality of epidermal tissue blisters into the at least one epidermal tissue harvesting and dissociation device.

[0020] According to another exemplary embodiment of the present disclosure, the at least one epidermal tissue harvesting and dissociation device is placed in at least one centrifuge rotor of the tissue processing instrument and tighten with a lid, the at least one tissue harvesting and dissociation device with the plurality of epidermal tissue blisters mixed by rotating the at least one centrifuge rotor in opposite directions that aids in tissue dissociation.

[0021] According to another exemplary embodiment of the present disclosure, the centrifuge rotor chamber designed to maintain the optimum temperature of 37°C for enzymatic dissociation of the tissue.

[0022] According to another exemplary embodiment of the present disclosure, the 70 micron cell strainer inside the tissue harvesting and dissociation device aids only single cells pass through towards the bottom of the device during dissociation process.

[0023] According to another exemplary embodiment of the present disclosure, the at least one epidermal tissue harvesting and dissociation device is centrifuged to settle individual cells at bottom of the at least one tissue processing instrument, an enzymatic solution is removed from the second port of the at least one epidermal tissue harvesting and dissociation device.

[0024] According to another exemplary embodiment of the present disclosure, a washing buffer is added to the at least one epidermal tissue harvesting and dissociation device through the second port and mixed by rotating the at least one centrifuge rotor forward and reverse directions,

[0025] According to another exemplary embodiment of the present disclosure, at final step, the individual cells are settled at the bottom of the at least one tissue processing instrument by centrifugation a single cell suspension is dissolved in the washing buffer and suspend into the solution.

[0026] According to another exemplary embodiment of the present disclosure, the individual cell suspension is collected through the second port into an application syringe and used for the transplant purpose.

BRIEF DESCRIPTION OF DRAWINGS
[0027] Other objects and advantages of the present invention will become apparent to those skilled in the art upon reading the following detailed description of the preferred embodiments, in conjunction with the accompanying drawings, wherein like reference numerals have been used to designate like elements, and wherein:

[0028] FIG. 1 is a diagram depicting an epidermal tissue harvesting and dissociation device of a point of care tissue processing system for epidermal tissue harvesting cum dissociation, according to an exemplary embodiment of the present disclosure.

[0029] FIG. 2 is a diagram depicting a tissue processing instrument of the point of care tissue processing system and its parts, according to an exemplary embodiment of the present disclosure.

[0030] FIG. 3 is a diagram depicting an epidermal tissue harvesting and dissociation device attachment to the Leg or hand of the person for epidermal tissue harvesting, according to an exemplary embodiment of the present disclosure.

[0031] FIG. 4 is a diagram depicting a second embodiment of the point of care tissue processing system connecting to epidermal harvesting and dissociation device through vacuum connector and skin warming component through other wired connectors, according to an embodiment of the present disclosure.

[0032] FIG. 5 is a diagram depicting a third embodiment of the point of care tissue processing system major internal parts along with epidermal tissue harvesting and dissociation device placement in the rotor of the system, according to an embodiment of the present disclosure.

[0033] FIG. 6 is a flowchart depicting an exemplary method for epidermal tissue harvesting cum dissociation, in accordance with one or more exemplary embodiments.

DETAILED DESCRIPTION
[0034] It is to be understood that the present disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.
[0035] The use of “including”, “comprising” or “having” and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. Further, the use of terms “first”, “second”, and “third”, and the like, herein do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.

[0036] Referring to FIG. 1 is a diagram 100 depicting the single use epidermal tissue harvesting and dissociation device of a point of care tissue processing system for epidermal tissue harvesting cum dissociation, according to an exemplary embodiment of the present disclosure. The epidermal tissue harvesting and dissociation device 100 may be used for epidermal tissue harvesting (by micro blister formation under vacuum and slicing) and dissociation to make a single cell suspension. The epidermal tissue harvesting and dissociation device 100 includes components such as a tissue harvesting component 101 and a dissociation component 103. The tissue harvesting component 101 may be a blister formation and slicing component. The dissociation component may be a part where tissue dissociation and separation of single cells from tissue occurs through cell strainer.

[0037] Tissue harvesting component 101 comprises of one upper plastic sheet 107 and one lower plastic sheet 105 that holds the three stainless steel (Hereafter called as SS) plates used for tissue blister slicing. The three SS sheets are 0.2mm thick and contain perforations 115 for epidermal tissue blister raising under vacuum conditions. The one middle SS slicing sheet 109 is sandwiched between upper SS sheet 113 and lower SS sheet 111. The Middle SS sheet 109 is protruded out of the device and connected to separate Lever 121 for tissue blister slicing by horizontal movement between upper SS sheet 113 and lower SS sheet 111. The perforations 115 may be closed by silicone cap 123 with protrusions after tissue slicing is completed.
[0038] The dissociation component 103 includes an oval shaped container 125 embedded with a cell strainer 127 for individual cells passing through after dissociation to bottom of the device. The cell strainer 127 may contain 70um pores for allowing only dissociated cells to pass through and retain the non-dissociated tissue above the cell strainer. The cell strainer 127 may contain strainer teeth 129 for aiding in tissue disintegration. The dissociation component 103 further includes a first port 131 and a second port 133. The first port 131 may be configured for vacuum creation and the second port 133 may be configured for solution addition/removal from the dissociation component 103. A “T” shaped tube 135 may be inserted in the dissociation component 103 and connected to individual ports 131, 133 in which one line opened to the bottom of the dissociation component 103 used for final cell product collection. One line of the “T” tube 135 may be opened just below the cell strainer 127 to create a vacuum and removal/addition of washing solution.

[0039] Referring to FIG. 2 is a diagram 200 depicting a tissue processing instrument of the point of care tissue processing system, according to an exemplary embodiment of the present disclosure. The tissue processing instrument 200 includes a control panel 202, a lid 204, an outer casing 206, a rotor chamber 208, a centrifuge rotor 210, a motor 212, a vacuum pump 214, a first Peltier module 216, a second Peltier module 218, a bottom support casing 220, and a skin warming component 222. The tissue processing instrument 200 may be a programmable centrifuge with the rotor chamber 208. The tissue processing instrument 200 may be fitted with the vacuum pump 214 and the skin warming component 222 that is used for micro blister creation in the epidermal tissue harvesting and dissociation device.

[0040] The centrifuge rotor 210 may be fixed to the motor 212 and placed inside the rotor chamber 208. The centrifugal rotor 210 may be configured to adapt the epidermal tissue harvesting and dissociation device 100 during tissue dissociation. The motor 212 may be a brushless DC motor for mixing and /or centrifuging the epidermal tissue harvesting and dissociation device during tissue dissociation. The temperature of rotor chamber 208 may be regulated by the Peltier modules 216, 218.The Peltier modules 216, 218 may be configured with a heat sink and a fan. The control panel 202 may be configured to control the vacuum pump 214, the temperature of rotor chamber 208, speed of the centrifuge rotor 210, temperature of the skin warming component 222. The speed of the centrifuge rotor 210 may be adjusted with a timer and speed controller in the control panel 202. The vacuum pump 214 and the skin warming component 222 may be configured to aid in tissue harvesting procedure.

[0041] The vacuum pump 214 may include vacuum capacity 80Kpa. The temperature controlled chamber 208 may be configured to maintain the required temperature for enzymatic tissue dissociation. The Peltier modules 216, 218 may be positioned under the temperature controlled chamber 208 aids in temperature maintenance in the temperature controlled chamber 208. The motor 212 may be controlled to mix the tissues for dissociation and centrifuge to collect the single cell suspension. The centrifuge rotor 210 may be fixed to the motor 212 is protruded to the temperature controlled chamber 208 that required for tissue device placement. The control panel 202 may be configured to control the motor 212 movement, the temperature controlled chamber 208, and the skin warming component 222. The control panel 202 may also be configured to program the processing of the tissue processing instrument. The components such as the temperature controlled chamber 208, the centrifuge rotor 210, the motor 212, the vacuum pump 214, the Peltier module 216, 218 may be placed in the outer casing 206 and the lid 204 made of powder coated metal.

[0042] Referring to FIG. 3 is a diagram 300 depicting a first embodiment of the point of care tissue processing system for epidermal tissue harvesting and dissociation device 100 attachment to the personnel leg/hand 307 with the band 305 and placement of skin warming component 222 for warming the skin during epidermal tissue blister formation and harvesting. The heating element may be used for temperature creation inside the skin warming component. The warming control sensor 301 is extended from skin warming component 222 and connecting to tissue processing instrument 200 for controlling the temperature of the skin warming component.

[0043] Referring to FIG. 4 is a diagram 400 depicting a second embodiment of the point of care complete tissue processing system 400 configuration and connections between epidermal tissue harvesting device 100 and tissue processing instrument 200, according to an embodiment of the present disclosure. The point of care tissue processing system 400 includes the epidermal tissue harvesting and dissociation device 100, the tissue processing instrument 200, the control panel 202, and the skin warming component 222, the warming control sensor connection 301, the vacuum connector 303, and the tightening band 305.

[0044] Referring to FIG. 5 is a diagram 500 depicting a third embodiment of the parts of the point of care tissue processing system for the epidermal tissue harvesting cum dissociation, according to an embodiment of the present disclosure. The point of care tissue processing system 500 showing the internal parts of tissue processing instrument and placement of epidermal tissue harvesting and dissociation device 100 in the centrifuge rotor 210, and the configuration of the motor 212, the vacuum pump 214, the first Peltier module 216, the second Peltier module 218, and the bottom support casing 220.

Tissue harvesting procedure:
[0045] The epidermal tissue harvesting and dissociation device 100 may be attached to a leg/hand of the person 307 and tightened with the elastic adjustable band 305.

[0046] The skin warming component 222 may be placed over the epidermal tissue harvesting and dissociation device 100 and observed the temperature to reach 45°C inside the skin warming component 222 with the control panel of the tissue processing instrument 202. The control panel 202 of the tissue processing instrument 200 may be configured to display the set and present temperature of the skin warming component 222. The first port 131 of the epidermal tissue harvesting and dissociation device 100 may be connected to the tissue processing instrument 200 through the vacuum connector 303 and the vacuum is applied at 65-70Kpa. The vacuum may be applied for 45-50 mins or 60 mins in certain cases until tissue blisters are raised into the perforations 115 of the tissue harvesting component 101. The raised epidermal tissue blisters may be sliced by moving the middle SS slicing sheet 109 horizontally with the help of the specific lever 121.The specific lever 121 may be the roller lever. The middle SS slicing sheet 109 may be configured to move horizontally in between the upper SS sheet 113 and lower SS sheet 111to cut the tissue blisters. The middle slicing sheet 109 may be moved back to its original place after the slicing of the tissue blisters is done. The tissue harvesting and dissociation device100 may be then detached from the leg/hand of the person 307 and placed on the table vertically. The perforations of the tissue slicing component 115 may be closed with the silicone cap 123 that contain protrusions to fill in the perforations 115.

Tissue Dissociation procedure:
[0047] 15ml of 0.5% Trypsin: ETHYLENEDIAMINETETRAACETIC ACID (EDTA) may be added to the first embodiment of the tissue dissociation device 100 for the tissue dissociation through the second port 133. The epidermal tissue harvesting and dissociation device 100 may be placed in the centrifuge rotor 210 of the tissue processing instrument 200.

[0048] The temperature of the rotor chamber 208 may be raised to 37°C which is optimum for enzymatic tissue dissociation. The motor 212 may be controlled to move forward and reverse to mix tissue for dissociation along with temperature maintenance in rotor chamber 208. The centrifuge rotor 210 in the rotor chamber 208 adapts the placement of Tissue harvesting and dissociation device 100.

[0049] The epidermal tissue harvesting and dissociation device 100 may be placed in the centrifuge rotor 210 of the tissue processing instrument 200 and tighten with the lid 204. The control panel 202 may be configured to control the movement of the motor 212 through which the rotor 210, the temperature of the rotor chamber 208.

[0050] The first embodiment of the tissue harvesting and dissociation device 100 with tissue pieces may be mixed for 40-60 min by rotating the centrifuge rotor 210 in opposite directions. The cell strainer 127 in the epidermal tissue harvesting and dissociation device 100 may be configured to allow dissociated cells to pass through the 70 micrometer pores to the bottom of the epidermal tissue harvesting and dissociation device 100 and non-dissociated tissue pieces may be retaining on top of the cell strainer 127. After a while (for example, 60 minutes),the epidermal tissue harvesting and dissociation device 100 may be centrifuged at 1200 rpm to 1500 rpm for 5 min to settle the individual cells at bottom of the tissue harvesting and dissociation device. After completion of dissociation, the enzymatic solution may be removed from the second port 133 and discarded.

[0051] Further 15- 20 ml of cell washing buffer may be added to the tissue harvesting and dissociation device 100 through the second port 133 and mixed for 5 minutes by rotating the centrifuge rotor 210 forward and reverse. Then cells may be settled in the bottom by centrifuging the device for 5 min at 1200 – 1500 rpm. The washing step may be repeated for removing any traces of the enzyme solution. Finally, single cell suspension is dissolved in 0.5 -1 ml of tissue wash buffer and suspend the cells into the solution. The individual cell suspension may be collected through the second port 133 into the application syringe.

[0052] The collected cell suspension into the syringe will be used for application onto the affected region of the skin surface.

[0053] Referring to FIG. 6 is a flowchart 600 depicting an exemplary method for epidermal tissue harvesting cum dissociation, in accordance with one or more exemplary embodiments. As an option, the method 600 is carried out in the context of the details of FIG. 1, FIG. 2 FIG. 3, FIG. 4, and FIG.5. However, the method 600 is carried out in any desired environment. Further, the aforementioned definitions are equally applied to the description below.

[0054] The method commences at step 602, attaches the epidermal tissue harvesting and dissociation device onto the leg/hand of the person and tightens with the band. Thereafter, at step 604, place the skin warming component over the epidermal tissue harvesting and dissociation device and the temperature inside the skin warming component may be adjusted in the control panel of the tissue processing instrument. Thereafter, at step 606, connect the vacuum port of the epidermal tissue harvesting and dissociation device to the tissue processing instrument through a tube connector and apply the vacuum at 65kpa. Thereafter, at step 608, observe for blister formation through the perforations and tissue blisters may be sliced by moving tissue slicing plates outwards with the help of the specific lever. Thereafter, at step 610, remove the epidermal tissue harvesting and dissociation device from the leg/hand and close the perforations with the silicon cap. Thereafter, at step 612, add the enzyme solution to the epidermal tissue harvesting and dissociation device and mix the enzyme solution properly. Thereafter, at step 614, place the epidermal tissue harvesting and dissociation device in the rotor of the tissue processing instrument and tighten with the lid.

[0055] Thereafter, at step 616, adjust the temperature of the rotor chamber and select the mix option through the control panel to mix tissues for tissue dissociation. Thereafter, at step 618, observe for the tissue dissociation and centrifuge for a certain time to settling cells at the bottom of the tissue dissociation device. Thereafter, at step 620, collect and discard the enzyme solution from the epidermal tissue harvesting and dissociation device. Thereafter, at step 622, add a washing solution (Phosphate Buffered Saline)/ (PBS) and mix the epidermal tissue harvesting and dissociation device for washing the cells and centrifuge to settle the cells at bottom of the epidermal tissue harvesting and dissociation device. Thereafter, at step 624, remove the washing solution and repeat the washing step. Thereafter, at step 626, observe the cells are precipitated at the bottom of the epidermal tissue harvesting and dissociation device. Thereafter, at step 628, mix the epidermal tissue harvesting and dissociation device in the instrument to suspend the precipitated cells into the solution. Thereafter, at step 630, collect the cell suspension through the cell collection port into the syringe.

[0056] Although the present disclosure has been described in terms of certain preferred embodiments and illustrations thereof, other embodiments and modifications to preferred embodiments may be possible that are within the principles and spirit of the invention. The above descriptions and figures are therefore to be regarded as illustrative and not restrictive.

[0057] Thus the scope of the present disclosure is defined by the appended claims and includes both combinations and sub-combinations of the various features described hereinabove as well as variations and modifications thereof, which would occur to persons skilled in the art upon reading the foregoing description.