The technique of the present disclosure relates to a filter for filtration, a container with a filter, and a method for removing foreign substances contained in a cell suspension.
Background technology
[0002]
　In recent years, cells have been collected from the body fluids and tissues of the patient or the donor, the collected cells have been cultured, and the obtained cells have been directly transplanted into the affected area, or the scaffolding material in which the cells have been seeded has been transplanted into the affected area. Attention is being paid to cell medicine and regenerative medicine that treat diseases in Japan. For some tissues such as skin, cornea, bone, and cartilage, cell medicine and regenerative medicine using these are actually performed, and are expected as next-generation treatment methods.
[0003]
　In order to perform cell medicine and regenerative medicine, it is necessary to culture and proliferate cells collected from patients to secure a certain number of cells. Cells that have grown to a certain number are collected from the culture medium, washed, and concentrated so that they can be used as cells for cell therapy and regenerative medicine.
[0004]
　However, since the cell suspension after culturing contains a medium, serum, a carrier for culturing, cell-derived waste products, debris, etc., it is necessary to separate and remove them. Further, it is necessary to wash and concentrate the cells even after removing them, and a centrifugal separation method is known as a method for performing these. For example, in International Publication No. 2013/114845, a culture container for culturing cells, a medium storage container for storing media and the like, a cell injection container for injecting cells, and a cell suspension after culturing. A cell culture kit has been proposed in which a cell collection container for collecting turbid liquid is connected by a conduit to construct a closed system environment. According to such a cell culture kit, it is possible to perform from cell injection to addition of medium, sampling, and recovery while maintaining a closed system in the kit.
[0005]
　In International Publication No. 2013/114845, when collecting the cultured cells, the culture vessel was allowed to stand to settle the cells in the cell culture solution, and then the supernatant of the cell culture solution was discharged to measure the amount of the solution. An example is shown in which the reduced and then concentrated cell culture medium is transferred from the culture vessel to the cell recovery vessel.
[0006]
　However, in order to recover the cells in this manner, the culture vessel must be allowed to stand to settle the cells in the cell suspension prior to discharging the supernatant of the cell culture solution, and the cells must be settled. It takes time to settle. Furthermore, even if the cells in the cell culture medium are precipitated over a sufficient period of time, the cells may be mixed in the supernatant by the discharge operation, and the cells may be discharged together with the supernatant.
[0007]
　International Publication No. 2014/007382 describes that sufficient welding strength was obtained by laser welding in which a molten resin layer was allowed to penetrate into the pores of a metal filter to weld each other to the opposing resin layers. There is. However, there is a problem that the metal filter is inflexible as used for cell treatment of a closed system.
[0008]
　Japanese Unexamined Patent Publication No. 2006-231875 describes a fluid filter device for vehicles in which a fiber (nonwoven fabric) made of polyester resin is used as a filter and the fiber (nonwoven fabric) is sandwiched between case constituent members. In Japanese Patent Application Laid-Open No. 2006-231875, one of the case constituent members is made of a resin such as nylon 66 or nylon 6 which is not colored with a pigment or the like, and the other is made of a resin such as nylon 66 or nylon 6 which is colored with a pigment or the like. It is stated that.
Outline of the invention
Problems to be solved by the invention
[0009]
　An object of the technique of the present disclosure is a filtration filter in which a filter is fixed between two polymer films with high welding strength, a filter-equipped container in which a filtration filter is welded so as to partition the inside, and a filter-equipped container. It is to provide a method for removing a foreign substance contained in a cell suspension using.
Means to solve problems
[0010]
　The inventors of the technique of the present disclosure prepare a filter for filtration in which a filter is fixed between two polymer films, and the material of the polymer film capable of obtaining high welding strength between these polymer films and the filter. And the film thickness thereof, and the opening rate of the filter were found, and the technique of the present disclosure was completed based on this finding.
[0011]
　That is, the technique of the present disclosure is composed of, for example, a flexible polymer film having a thickness of 120 μm or more containing a polymer, and is formed in a frame shape having a first passage hole penetrated in the thickness direction inside. A second welding frame composed of a first welding frame and a flexible polymer film having a thickness of 120 μm or more containing a polymer, and formed in a frame shape having a second passage hole penetrated in the thickness direction inside. The frame portion of the first welding frame is made of a material having a melting point higher than that of the material of these polymer films, having pores, and having a pore opening rate of 10% or more and 80% or less, and the outer peripheral portion is the frame portion of the first welding frame. The first welding frame includes a filter welded to the first welding frame and the second welding frame in a state of being sandwiched between the entire circumference of the first welding frame and the entire circumference of the frame portion of the second welding frame. , High density polyethylene (HDPE: High Density Polyethylene) having a melting point of 120 ° C. to 140 ° C., linear low density polyethylene (LLDPE: Linear Low Density Polyethylene) having a melting point of 105 ° C. to 125 ° C., or the high density polyethylene and A filter for filtering, which is composed of a polymer containing a mixture of at least one of the linear low-density polyethylenes.
[0012]
　Here, the "opening rate" refers to the ratio of the area of ​​the hole to the total area.
[0013]
　The technique of the present disclosure also includes a container with a filter, wherein the filter for filtration is welded inside a container made of a polymer, thereby partitioning the inside of the container.
[0014]
　Further, one embodiment of the technique of the present disclosure is to inject a cell suspension containing or may contain foreign matter into one compartment inside the container in such a container with a filter, and filter the cell suspension for filtration. The present invention relates to a method for removing foreign substances contained in a cell suspension by collecting a filtrate containing cells from the other compartment. The filter has a pore that communicates in the thickness direction, and the pore has a pore diameter that allows cells to pass through, but foreign matter of a certain size or larger cannot pass through. Therefore, such foreign matter can be removed from the cell suspension by passing it through a filter for filtration.
[0015]
　Further, one embodiment of the technique of the present disclosure is a filtrate containing cells by injecting a cell suspension into one compartment inside the container in such a filter-equipped container and passing the cell suspension through a filter for filtration. Is further collected, a solution for cell suspension is injected into the same compartment, the cells remaining in the same compartment are resuspended, and the resuspended cell suspension is passed through a filter for filtration to obtain cells. The present invention relates to a method for removing foreign substances contained in a cell suspension, which comprises a step of collecting a filtrate containing the above.
The invention's effect
[0016]
　In the filtration filter and the container with a filter of the technique of the present disclosure, the portion of the filter that is not sandwiched between the first welding frame and the second welding frame, that is, the portion corresponding to the first passing hole and the second passing hole is the filter. Demonstrate the function as. Here, since the filter and the first welding frame and the second welding frame are welded with high welding strength, the welded portion is hard to peel off and there is little possibility that the fluid leaks from the welded portion.
[0017]
　Therefore, even when the method of removing foreign substances contained in the cell suspension of the technique of the present disclosure using the container with a filter of the technique of the present disclosure is carried out, there is little risk of liquid leakage from the welded portion.
A brief description of the drawing
[0018]
FIG. 1 is a plan view showing a container with a filter according to an embodiment.
FIG. 2 is an exploded perspective view showing a container with a filter according to an embodiment.
FIG. 3 is a schematic perspective view showing a container with a filter according to an embodiment.
FIG. 4 is a cross-sectional view taken along the line AA in FIG.
Mode for carrying out the invention
[0019]
　As illustrated in FIG. 2, one embodiment of the technique of the present disclosure is higher than the material of the first welding frame 10 and the second welding frame 12 between the first welding frame 10 and the second welding frame 12. A filtration filter 16 in which a flat filter 14 is fixed by welding, which is made of a material having a melting point and has a portion not sandwiched between the first welding frame 10 and the second welding frame 12.
[0020]
　Hereinafter, each member constituting the filtration filter in the technique of the present disclosure will be described in detail by taking the filtration filter 16 shown in FIG. 2 as an example. The material of the filter 14 is polyolefin such as polypropylene and polyethylene, polyester, polyvinyl chloride, polyvinyl alcohol, vinylidene chloride, acrylic polymer such as polymethylmethacrylate and polyacrylonitrile, polyamide such as nylon, polystyrene, polyurethane, polyimide, aramid, etc. It preferably contains at least one selected from the group consisting of polyether ether ketones, polysulfones, rayons, celluloses, chitins, chitosans, cotton, hemp, glass, carbon fibers, and metals. Among them, it is preferable to contain at least one selected from the group consisting of polyester, polyamide, polyolefin, polyether ether ketone, polyether sulfone, carbon fiber, and metal, and preferably contains polyester, polyamide, polyolefin, polyether ether ketone, and poly. It is more preferably at least one selected from the group consisting of ether sulfone, carbon fiber, and metal, most preferably polyester or polyamide.
[0021]
　Here, specific examples of polyester include polyethylene terephthalate, polybutylene terephthalate, polymethylene terephthalate and the like. Specific examples of the polyamide include 6,6-nylon, 6-nylon, 12-nylon and the like.
[0022]
　The material of the filter 14 may be a porous body shape having a communicating hole structure having a plurality of pores 14A, an aggregate of fibers, a non-woven fabric, a woven fabric, a knitted fabric, or the like, but is preferably a woven fabric or a knitted fabric.
[0023]
　The pore diameter of the pore portion 14A of the filter 14 is preferably 5 μm or more and 200 μm or less, which is a diameter required for capturing impurities such as culture carrier residue other than cells. If the pore diameter of the pore portion 14A is smaller than 5 μm, the filter 14 may be clogged and the efficiency of removing contaminants may decrease. On the other hand, if the pore size of the pore 14A is larger than 200 μm, it becomes difficult to capture impurities and target cells. The pore size of the pore 14A is preferably 10 μm to 200 μm from the viewpoint of the efficiency of removing contaminants such as a relatively large culture carrier residue and the ability to capture target cells.
[0024]
　The opening ratio of the filter 14 is 10% or more and 80% or less in view of the welding strength. If the pore size is lower than 10%, the melted polymer film is less likely to be entangled with the filter 14 when welded to the polymer films constituting the first welding frame 10 and the second welding frame 12, causing problems such as delamination. It is easy to cause. On the other hand, if the opening rate of the filter 14 is larger than 80%, the strength of the filter 14 may decrease, which may cause cutting, cracking, or cracking, and the mechanical strength of the filter 16 for filtration may decrease. There is. The opening rate of the filter 14 is preferably 10% or more and 70% or less, and more preferably 10% or more and 50% or less.
[0025]
　The material of the filter 14 has a higher melting point than the materials of the first welding frame 10 and the second welding frame 12. The material of the filter 14 is preferably one having a high melting point in the range of 80 ° C. or higher and 180 ° C. or lower as compared with the melting points of the materials of the first welding frame 10 and the second welding frame 12, preferably 100 ° C. or higher. Those having a high melting point in the range of 160 ° C. or lower are more preferable, and those having a high melting point in the range of 110 ° C. or higher and 150 ° C. or lower are further preferable. For example, when the materials of the first welding frame 10 and the second welding frame 12 are high-density polyethylene having a melting point of 120 ° C. to 140 ° C., the melting point of the material of the filter 14 is preferably 200 ° C. or higher, 320. ° C. or lower, more preferably 220 ° C. or higher and 300 ° C. or lower, and even more preferably 230 ° C. or higher and 290 ° C. or lower. Further, for example, when the material of the first welding frame 10 and the second welding frame 12 is linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., the melting point of the material of the filter 14 is preferably 185 ° C. or higher. It is 305 ° C. or lower, more preferably 205 ° C. or higher and 285 ° C. or lower, and further preferably 215 ° C. or higher and 275 ° C. or lower.
[0026]
　The film thickness of the filter 14 is preferably 50 μm or more and 200 μm or less from the viewpoint of the strength of the filter 14 after welding. If the film thickness of the filter 14 is smaller than 50 μm, the strength of the filter 14 may decrease, which may cause cutting, cracking, or cracking, and the mechanical strength of the filter 16 for filtration may also decrease. When the film thickness of the filter 14 is larger than 200 μm, the pores of the filter 14 in the region close to the surface of the filter 14 when the polymer films constituting the first welding frame 10 and the second welding frame 12 are melted. The molten polymer film material permeates only in 14A, and welding of the first welding frame 10 and the second welding frame 12 facing each other is less likely to occur, and the welding strength may be insufficient.
[0027]
　The materials of the first welding frame 10 and the second welding frame 12 are polypropylene, polyethylene, high-density polyethylene, low-density polyethylene, linear low-density polyethylene, polyolefins such as ethylene-propylene copolymer, polyester, polyvinyl chloride, and polyvinyl. Alcohol, vinylidene chloride, polystyrene, acrylic polymers such as polymethylmethacrylate and polyacrylonitrile, polyamide, polyurethane, polyimide, aramid, polyether ether ketone, polysulfone, and a mixture of one or more selected from the group consisting of carbon. It is preferable to have. Among them, a mixture of one or more selected from the group consisting of polyolefin polyesters such as polypropylene, polyethylene, high-density polyethylene, low-density polyethylene, linear low-density polyethylene, polyamide, and polyvinyl alcohol is more preferable, and high-density polyethylene or a mixture of a plurality of them is more preferable. Most preferably, it is linear low density polyethylene. In particular, when the material is high-density polyethylene, the melting point is preferably 120 ° C. to 140 ° C., and when the material is linear low-density polyethylene, the melting point is preferably 105 ° C. to 125 ° C.
[0028]
　Here, the combination of the mixture of two or more kinds of materials is not particularly limited, but the combination of polyethylene and polyamide, polyethylene and polyvinyl alcohol is particularly preferable.
[0029]
　The density of the high-density polyethylene having a melting point of 120 ° C. to 140 ° C. used as the material of the first welding frame 10 and the second welding frame 12 is approximately 910 kg / m 3 to 950 kg / m 3 , and the melting point is 105. The density of the linear low-density polyethylene at ° C. to 125 ° C. is approximately 912 kg / m 3 to 930 kg / m 3 .
[0030]
　The melting point of the above-mentioned material is obtained as the temperature at the heat absorption peak position in the measurement using a differential scanning calorimetry (DSC), and the density of the above-mentioned material is JIS K 0061. It is obtained by measuring by the in-liquid weighing method described in "Method for measuring density and specific gravity of chemical products" and JIS Z 8807 "Method for measuring density and specific gravity of solids".
[0031]
　In the filtration filter 16 of the technique of the present disclosure, both the first welding frame 10 and the second welding frame 12 have a frame shape having a first passing hole 10A or a second passing hole 12A penetrating in the thickness direction inside. And
i) First passage hole 10A in the frame of the first welding frame 10
ii) Of the filter 14, it is in contact with the frame portions 10B and 12B of the first welding frame 10 and the second welding frame 12, respectively. No portion
iii) The
first welding frame 10, the filter 14, and the first welding frame 10, the filter 14, and the second 2 It is preferable that the welding frame 12 is welded.
[0032]
　As described above, when both the first welding frame 10 and the second welding frame 12 have a frame shape, they may have the same shape or different shapes, but at the punched portion of the first welding frame 10. It is preferable that a certain first passage hole 10A and the second passage hole 12A which is a punched portion of the second welding frame 12 have the same shape, and among them, those having a rectangular shape (including a square) are preferable. It is preferable that the first welding frame 10 and the second welding frame 12 both have a shape similar to the outer shape of the first welding frame 10 and the second welding frame 12 (in this case, the widths of the frame portions 10B and 12B are substantially uniform). Will be). In particular, the first passage hole 10A of the first welding frame 10 and the second passage hole 12A of the second welding frame 12 have the same rectangular shape, and the outer shape of the first welding frame 10 and the second welding frame 12 It is preferable that the outer shapes are all similar to each other in a rectangular shape, and in particular, the outer shape of the first welding frame 10 and the outer shape of the second welding frame 12 are the same rectangular shape.
[0033]
　In the planar filter 14 used in the technique of the present disclosure, it is the portion of the first welding frame 10 that is not sandwiched between the first welding frame 10 and the second welding frame 12 that functions as the filter 14. Although it is a portion of the passing hole 10A and the second welding frame 12 corresponding to the second passing hole 12A, the first welding frame 10 and the second welding frame 12 are both frame-shaped in the filtering filter 16 of the technique of the present disclosure. In some cases, as described above, the fluid in the object to be filtered is the first passage hole 10A in the frame of the first welding frame 10, the first passage hole 10A of the first welding frame 10 in the filter 14, and the second welding frame. The portion of the 12 corresponding to the second through hole 12A moves in the order of the second through hole 12A in the frame of the second welding frame 12, or vice versa, while the first welding frame 10 and the second welding frame 12 In the part where is welded, the movement of such fluid is blocked.
[0034]
　Therefore, when the first welding frame 10 and the second welding frame 12 are both frame-shaped in the filtration filter 16 of the present disclosure technique, the filter 14 in the filtration filter 16 of the present disclosure technique exerts its function. The outer shape of the filter 14 is the same as that of the first welding frame 10 and the second welding frame 12 on the entire circumference of the first passing hole 10A of the first welding frame 10 and the second passing hole 12A of the second welding frame 12. It must be able to form an overlap.
[0035]
　The shape of the filter 14 used in the technique of the present disclosure is preferably rectangular (including a square), in which case the first welding frame 10 and the second welding frame 12 are both of the filter 14 having such a shape. It is preferable that they have the same shape that overlaps at the peripheral edges.
[0036]
　The film thickness of the first welding frame 10 and the second welding frame 12 is 120 μm or more because it is necessary to obtain high welding strength with the filter 14. The upper limit of the film thickness of the first welding frame 10 and the second welding frame 12 is not particularly limited from the viewpoint of the welding strength with the filter 14, but it is preferably 500 μm or less from the viewpoint of providing flexibility. The film thickness of the first welding frame 10 and the second welding frame 12 is more preferably 200 μm or more and 400 μm or less, and most preferably 200 μm or more and 300 μm or less.
[0037]
　The welding strength of the welded portion 34 is preferably 20 N / 15 mm or more, more preferably 23 N / 15 mm or more, still more preferably 23 N / 15 mm or more, when measured by the method described in Example B (method for evaluating the welding strength). It is 30 N / 15 mm or more. For example, the welding strength of the welded portion 34 is 20N / 15mm to 80N / 15mm, 23N / 15mm to 60N / 15mm, or 30N / 15mm to 60N / 15mm. The standard value of JIS Z0238 is 23 N / 15 mm, but even in the filtration filter 16 of the technique of the present disclosure, this value serves as a guideline for the welding strength of the welded portion 34 to be achieved. However, even if the welding strength of the welded portion 34 falls below this value, it does not mean that it cannot be used immediately.
[0038]
　The filtering filter 16 in the technique of the present disclosure is a filter 14 sandwiched between a first welding frame 10 and a second welding frame 12, and the filters 14 are placed at a temperature equal to or higher than the melting point of the first welding frame 10 or the second welding frame 12. By heating at a temperature equal to or lower than the melting point of the filter 14, the molten polymers of the first welding frame 10 and the second welding frame 12 are allowed to penetrate into the pores 14A of the unmelted filter 14, and then, for example, heat is dissipated. This refers to a laminate including three layers in which each of the first welding frame 10 and the second welding frame 12 and the filter 14 are fixed via a welding portion 34 by solidifying the polymer. In particular, the polymers melted from both sides of the filter 14 are connected through the pores 14A of the unmelted filter 14, and the first welding frame 10 and the second welding frame 12 facing each other are fixed by being bonded to each other. It is preferable that it is.
[0039]
　Then, the filtration filter 16 of the technique of the present disclosure may be welded to another member via at least one of the first welding frame 10 and the second welding frame 12. In this case, at least one of the first welding frame 10 and the second welding frame 12 in the technique of the present disclosure functions as a medium for realizing welding with other members.
[0040]
　As illustrated in FIG. 4, in one embodiment of the technique of the present disclosure, the filtration filter 16 is welded into a container 20 containing two polymer sheets (20A, 20B), thereby forming the inside of the container 20. Is a container with a filter 22 in which is divided. Hereinafter, the filter-equipped container in the technique of the present disclosure will be described in detail by taking the filter-equipped container 22 shown in FIG. 4 as an example.
[0041]
　Here, “partitioned” means, as shown in FIG. 4, from the section S1 which is a space composed of one surface of the filtration filter 16 and the inside of the container 20, the other of the filtration filter 16 The filtration filter 16 allows the container 20 to have a flow path for the fluid to move to the compartment S2, which is a space composed of the surface and the inside of the container 20, except for the filter 14 of the filtration filter 16. It means that the inside is partitioned. With such a configuration, the container with a filter 22 of the technique of the present disclosure functions as a filter.
[0042]
　The container 20 in the technique of the present disclosure includes two polymer sheets (20A, 20B). The material of these polymer sheets is flexible plastic, and specific examples thereof include the same ones as described above for the second welding frame 12 in the technique of the present disclosure, and preferred ones have the same physical properties (particularly melting point). However, it is particularly preferable that the material is compatible with the material of the first welding frame 10 and the material of the second welding frame 12. This is because a welding method can be used when fixing the filtration filter 16 of the technique of the present disclosure inside the container 20. For example, when the material of the first welding frame, the second welding frame and the flexible plastic is high-density polyethylene having a melting point of 120 ° C. to 140 ° C., the material of the flexible plastic has the same height. High density polyethylene can be preferably used. Further, for example, when the material of the first welding frame and the second welding frame is linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., the same linear low-density polyethylene can be used as the material of the flexible plastic. Polyethylene can be preferably used.
[0043]
　As the shape of the container 20, for example, two polymer sheets having the same shape, for example, having a rectangular shape are preferably opposed to each other and manufactured by welding the peripheral edges thereof.
[0044]
　In that case, the filter 16 for filtration is also made to have a substantially similar shape slightly smaller than the rectangular shape of the container 20 (see FIG. 1), and the first welding frame 10 is welded to one inner surface of the container 20 at the first welding portion 24. , It is preferable that the second welding frame 12 is welded to the other inner surface of the container 20 by the second welding portion 26. The polymer "sheet" constituting the container 20 and the polymer "film" constituting the first welding frame 10 and the second welding frame 12 are substantially the same materials having flexibility. In this embodiment, in order to make it easier to understand the parts used, "sheet" and "film" will be described separately.
[0045]
　As shown in FIG. 2, the first welding portion 24 is attached to one polymer sheet of the container 20 in the thickness direction of the first welding frame 10 (in the present embodiment, one end in the longitudinal direction of the container 20). The portion side) is formed in a substantially U shape so as to be open. Further, in the second welding portion 26, the other (in the present embodiment, the other end side in the longitudinal direction in the container 20) is opened to the other polymer sheet of the container 20 in the thickness direction of the second welding frame 12. It is formed in a substantially U shape. That is, the open portions of the first welded portion 24 and the second welded portion 26 are arranged on opposite sides.
[0046]
　Further, as shown in FIG. 3, the container 20 used in the technique of the present disclosure preferably includes at least one of an injection port 30 and a pouring port 32, and is used for filtration according to the technique of the present disclosure. It is particularly preferable that one of the compartments in the container 20 partitioned by the filter 16 is provided with the injection port 30 and the other compartment is provided with the pouring port 32. Here, the number of the injection port 30 and the ejection port 32 may be a plurality, and for example, the injection port 30 and the ejection port 32 may be provided in each of both compartments. Here, the injection port has a passage connecting the inside and the outside of the container, and is for injecting the cell suspension into the container 20. There is no other passage through which the cell suspension can be injected into the container 20 of the filter-equipped container 22. On the contrary, the injection port is for injecting the cell suspension from the inside of the container 20. There is no other passage through which the cell suspension can be poured into the container 20 of the filter-equipped container 22. That is, the polymer sheet 20A and the polymer sheet 20B are welded to each other in the open portions of the first welding portion 24 and the second welding portion 26, except for the locations where the injection port and the injection port are attached. There is. Further, the injection port and the injection port are welded or adhered to the polymer sheet 20A and the polymer sheet 20B, and the inside of the container 20 is sealed except for the passage provided by each port.
[0047]
　Further, the filter-equipped container 22 of the technique of the present disclosure may be one in which the inside of the container 20 is partitioned into a space of n + 1 by n (n is a natural number of 2 or more) filtration filters 16. In this case, a part or all of each section inside the container 20 may be provided with at least one of an injection port 30 and a discharge port 32 that can be connected to the inside of each section.
[0048]
　Further, as shown in FIG. 4, the technique of the present disclosure injects a cell suspension (not shown) into one compartment S1 inside the container 20 in the container with a filter 22 of the technology of the present disclosure, and from the other compartment S2. A method of removing foreign matter contained in a cell suspension, which comprises the step of recovering the cell suspension from which the foreign matter has been removed. Here, foreign substances such as unnecessary culture carriers and cell aggregates remain in the compartment S1 into which the cell suspension is injected.
[0049]
　Further, in the technique of the present disclosure, the cell suspension is injected into one compartment S1 inside the container 20 in the filter-equipped container 22 of the present disclosure technique, the cells are filtered by the filter 14, and the cells are resuspended. A method for removing foreign substances contained in a cell suspension, which comprises a step of injecting a liquid for use into the compartment S1, resuspending the cells, and recovering the cell suspension from the compartment S1.
[0050]
　In the method of removing foreign substances in these cell suspensions and the method of removing foreign substances contained in cells in cell suspensions, a separation operation of cell suspension components by a filter 14 is indispensable, and the driving force for this is essential. As the force, for example, a method such as the weight of the cell suspension itself, the pressurization of the compartment S1 inside the container 20 in which the cell suspension is injected, or the depressurization of the other compartment S2 can be used.
Example
[0051]
A. It has a method for measuring the melting point, density, and film thickness of the polymer film used as the first polymer film constituting the first welding frame 10 and the second polymer film forming the second welding frame 12, and the pores 14A. The method for measuring the pore size, the pore size, and the fiber diameter of the filter used as the filter 14 is as follows.
[0052]
(A-1) Method for Measuring Melting Point (Tm) of
　Polymer Film The melting point of the polymer film was measured by DSC (manufactured by TA Instruments, Q20). The DSC measurement conditions were a nitrogen atmosphere (50 ml / min), a measurement temperature range of 30 ° C. to 200 ° C., and a heating rate of 10 ° C./min. As the melting point of the polymer film, the temperature at the peak position of the endothermic peak observed in melting at DSC was used.
[0053]
(A-2) Method for Measuring Density of Polymer Film The density of the
　polymer film was measured by an in-liquid weighing method. In the in-liquid weighing method, a specific gravity measuring jig (manufactured by Mettler Toledo) was attached to a balance (manufactured by Mettler Toledo, Balance XS105), the polymer film was weighed in air, and then weighed in ethanol. The liquid temperature was measured, the density of ethanol was determined by the method described in the literature (Dlight E. Gray, American Institute of Physics Handbook, McGraw-Hill Book Company Inc., 1957), and the density of ethanol was determined by the following formula. Calculated.
　　　　　　ρ = {A / (AB)} × (ρ0-d) + d
[0054]
　Here, ρ is the density of the sample, A is the weight in the air, B is the weight in the liquid, ρ0 is the density of the liquid, and d is the density of the air (0.002 g / cm 3 ). show.
[0055]
(A-3) Method for measuring film thickness of polymer film For the film thickness of the
　polymer film, the catalog value presented by the film manufacturer was used.
[0056]
(A-4) Method for measuring the opening rate of the
　filter For the opening rate of the filter, the catalog value presented by the filter manufacturer was used.
[0057]
(A-5) Method for Measuring Hole Diameter of Filter Hole For the hole diameter of the
　filter, the catalog value presented by the filter manufacturer was used.
[0058]
(A-6) Method for measuring the fiber diameter of the
　filter For the fiber diameter of the filter, the catalog value presented by the filter manufacturer was used.
[0059]
B. Method for manufacturing filter 16 for filtration
　The outer circumference of the filter 14 is sandwiched between the first polymer film constituting the first welding frame 10 and the second polymer film constituting the second welding frame 12 over the entire circumference. In this state, the first welding frame 10 is heated by using an impulse sealer (SURE NL-102JW manufactured by Ishizaki Electric Co., Ltd.) set at 230 ° C., which is equal to or higher than the melting point of the polyethylene film and lower than the melting point of the PET filter 14. , The filter 14 and the second welding frame 12 were welded through the welding portion 34. The welding portion 34 was formed over the entire circumference of the portion sandwiched between the first welding frame 10 and the second welding frame 12 of the filter 14. In the filtration filter 16 produced in this way, the portion of the filter that is not sandwiched between the first welding frame 10 and the second welding frame 12, that is, the portion corresponding to the first passing hole 10A and the second passing hole 12A is used as a filter. Demonstrate function. Here, in the filtration filter 16, the materials of the first polymer film constituting the first welding frame 10, the second polymer film constituting the second welding frame 12, and the filter 14 are variously changed, and the following Example 1 The materials of the materials described in 18 to 18 and Comparative Examples 1 to 18 were produced. In Comparative Examples 19 to 20, the first polymer film and the second polymer film constituting the second welding frame 12 are welded to each other, which does not correspond to the filter 16 for filtration, but in the same manner. Welding strength was evaluated.
[0060]
C. Method
　for Measuring Welding Strength The welding strength of the welding portion 34 in the filtration filter 16 was measured using an evaluation model described in detail below.
　The welding strength was measured by a peeling test with reference to JIS Z0238. Three welded portions 34 of the filtration filter 16 welded by the impulse sealer were cut out in a size of 25 mm in length and 15 mm in width. This was subjected to a 180 ° peeling test method using a tensile tester (Ez-Test-Ez-SX, manufactured by Shimadzu Corporation). Here, the tensile tester set the tensile speed to 10 mm / sec and the inter-chuck distance to 20 mm / sec. The welding strength was determined as the maximum load until peeling or breaking of the welded portion 34 (N / 15 mm), and was evaluated as an average value of N = 3.
[0061]
　The standard value of JIS Z0238 is 23 N / 15 mm, but even in the filtration filter 16 of the technique of the present disclosure, this value serves as a guideline for the welding strength of the welded portion 34 to be achieved.
[0062]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET (polyethylene terephthalate) between two films. Filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C, length 80 mm, width 15 mm) is sandwiched and welded in this state for filtration. Filter 16 was obtained. The welding strength of the welded portion 34 was 32.6 N / 15 mm.
[0063]

　High-density polyethylene film (HDPE, POLYELITE EH manufactured by Hiroyuki Hosokawa, Tm = 126 ° C., density 0.947 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) made of PET between two sheets. A filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to filter for filtration. 16 was obtained. The welding strength of the welded portion 34 was 33.3 N / 15 mm.
[0064]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = (254 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 30.5 N / 15 mm.
[0065]

　Two linear low-density polyethylene films (LLDPE, Toyobo L4102, Tm = 123 ° C., density 0.907 g / cm 3 , film thickness 100 μm, L4102, length 80 mm, width 15 mm) are laminated and welded. Then, a film having a film thickness of 200 μm was prepared. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = (254 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 33.7 N / 15 mm.
[0066]

　Two linear low-density polyethylene films (LLDPE, Tamapoli SE620L, Tm = 113 ° C., density 0.921 g / cm 3 , film thickness 140 μm, SE620L, length 80 mm, width 15 mm) are laminated and welded. Then, a film having a film thickness of 280 μm was prepared. A PET filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254) between two linear low-density polyethylene films having a film thickness of 280 μm. (° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 26.7 N / 15 mm.
[0067]
PET between two
　linear low-density polyethylene films (LLDPE, Tamapoli SE620L, Tm = 113 ° C., density 0.921 g / cm 3 , film thickness 140 μm, SE620L, length 80 mm, width 15 mm) Filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C, length 80 mm, width 15 mm) is sandwiched and welded in this state for filtration. Filter 16 was obtained. The welding strength of the welded portion 34 was 20.2 N / 15 mm.
[0068]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) A filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to filter for filtration. 16 was obtained. The welding strength of the welded portion 34 was 13.9 N / 15 mm.
[0069]
Made of PET between two
　low-density polyethylene films (LDPE, POLYELITE EL manufactured by Hiroyuki Hosokawa, Tm = 115 ° C., density 0.907 g / cm 3 , film thickness 250 μm, length 80 mm, width 15 mm). A filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C., length 28 mm, width 15 mm) is sandwiched and welded in this state to filter for filtration. 16 was obtained. The welding strength of the welded portion 34 was 13.9 N / 15 mm.
[0070]
PET filter 14 (
　LDPE, manufactured by Sampler Tech, Tm = 110 ° C., density 0.915 g / cm 3 , film thickness 300 μm, length 80 mm, width 15 mm) NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254 ° C, length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. rice field. The welding strength of the welded portion 34 was 14.9 N / 15 mm.
[0071]
PET filter 14 (
　LDPE, manufactured by Sampler Tech, Tm = 111 ° C, density 0.915 g / cm 3 , film thickness 500 μm, length 80 mm, width 15 mm) NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254 ° C, length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. rice field. The welding strength of the welded portion 34 was 14.4 N / 15 mm.
[0072]
Between two
　linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) A PET filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C., length 80 mm, width 15 mm) was sandwiched between them and welded in this state. A filtration filter 16 was obtained. The welding strength of the welded portion 34 was 12.3 N / 15 mm.
[0073]
Made of PET between two
　linear low-density polyethylene films (LLDPE, Toyobo L4102, Tm = 123 ° C., density 0.907 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm). A filter 14 (NBC Meshtec No. T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore opening rate 23%, Tm = 254 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to filter for filtration. 16 was obtained. The welding strength of the welded portion 34 was 16.0 N / 15 mm.
　The results of Examples 1 to 6 and Comparative Examples 1 to 6 are summarized in Table 1.
[0074]
[table 1]

[0075]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) Nylon filter 14 between two films (03-30 / 18, manufactured by Cefar, pore diameter 30 μm, fiber diameter 40 μm, pore size 18%, Tm = 252 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 29.0 N / 15 mm.
[0076]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Nylon filter 14 (Sefar 03-30/18, pore diameter 30 μm, fiber diameter 40 μm, pore size 18%, Tm = 252 ° C.) between two linear low-density polyethylene films having a film thickness of 200 μm. (Length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 23.4 N / 15 mm.
[0077]
Made of nylon between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) A filter 14 (03-30 / 18, manufactured by Cefar, pore diameter 30 μm, fiber diameter 40 μm, pore size 18%, Tm = 252 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to form a filter 16 for filtration. Obtained. The welding strength of the welded portion 34 was 12.0 N / 15 mm.
　The results of Examples 7 and 8 and Comparative Example 7 are summarized in Table 2.
[0078]
[Table 2]

[0079]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (PEP24 manufactured by Cefar, pore diameter 21 μm, fiber diameter 41 μm, pore size 12%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 24.2 N / 15 mm.
[0080]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (PET 24 made by Safer, pore diameter 21 μm, fiber diameter 41 μm, pore size 12%, Tm = 257 ° C., length 80 mm). , Width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 24.2 N / 15 mm.
[0081]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (Sefar 07-27 / 19, pore diameter 27 μm, fiber diameter 35 μm, pore size 19%, Tm = 256 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 32.7 N / 15 mm.
[0082]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (Sefar 07-27 / 19, pore diameter 27 μm, fiber diameter 35 μm, pore size 19%, Tm = 256 ° C. (Length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 29.4 N / 15 mm.
[0083]

　High-density polyethylene film (HDPE, Tamapoli HD, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (T-380T manufactured by NBC Meshtec, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 32.6 N / 15 mm.
[0084]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (NBC Meshtec T-380T, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254 ° C. (Length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 30.5 N / 15 mm.
[0085]

　High-density polyethylene film (HDPE, Tamapoli HD, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (T-180T manufactured by NBC Meshtec, pore diameter 86 μm, fiber diameter 55 μm, pore size 37%, Tm = 255 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 52.2 N / 15 mm.
[0086]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (NBC Meshtec T-180T, pore diameter 86 μm, fiber diameter 55 μm, pore size 37%, Tm = 255 ° C. (Length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 35.8 N / 15 mm.
[0087]

　High-density polyethylene film (HDPE, Tamapoli HD, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (T-100T manufactured by NBC Meshtec, pore diameter 183 μm, fiber diameter 71 μm, pore size 52%, Tm = 255 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 53.6 N / 15 mm.
[0088]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (NBC Meshtec T-100T, pore diameter 183 μm, fiber diameter 71 μm, pore size 52%, Tm = 255 ° C. (Length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 35.6 N / 15 mm.
[0089]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (PET6-HD manufactured by Cefar, pore diameter 6 μm, fiber diameter 34 μm, pore size 5%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. The welding strength of the welded portion 34 was 13.1 N / 15 mm.
[0090]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between the two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (PET6-HD manufactured by Cefar, pore diameter 6 μm, fiber diameter 34 μm, pore size 5%, Tm = 257 ° C., length). A filter 16 for filtration was obtained by sandwiching (80 mm in width and 15 mm in width) and welding in this state. The welding strength of the welded portion 34 was 12.3 N / 15 mm.
[0091]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (PETE6-HD manufactured by Cefar, pore diameter 6 μm, fiber diameter 34 μm, pore size 5%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. .. The welding strength of the welded portion 34 was 6.0 N / 15 mm.
[0092]

　High-density polyethylene film (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) PET filter 14 (PEP15 manufactured by Cefar, pore diameter 15 μm, fiber diameter 37 μm, pore size 9%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. The welding strength of the welded portion 34 was 18.5 N / 15 mm.
[0093]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. Between two linear low-density polyethylene films having a thickness of 200 μm in this way, a PET filter 14 (PET15 made by Safer, pore diameter 15 μm, fiber diameter 37 μm, pore size 9%, Tm = 257 ° C., length 80 mm). , Width 15 mm) was sandwiched and welded in this state to obtain a filtration filter 16. The welding strength of the welded portion 34 was 17.1 N / 15 mm.
[0094]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (PEP15 manufactured by Cefar, pore diameter 15 μm, fiber diameter 37 μm, pore size 9%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. The welding strength of the welded portion 34 was 8.1 N / 15 mm.
[0095]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (PEP24 manufactured by Cefar, pore diameter 21 μm, fiber diameter 41 μm, pore size 12%, Tm = 257 ° C., length 80 mm, width 15 mm) was sandwiched and welded in this state to obtain a filter 16 for filtration. The welding strength of the welded portion 34 was 11.1 N / 15 mm.
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (Sefar 07-27 / 19, pore diameter 27 μm, fiber diameter 35 μm, pore opening rate 19%, Tm = 256 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to form a filter 16 for filtration. Obtained. The welding strength of the welded portion 34 was 13.8 N / 15 mm.
[0096]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (T-380T manufactured by NBC Meshtec, pore diameter 28 μm, fiber diameter 35 μm, pore size 23%, Tm = 254 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to form a filter 16 for filtration. Obtained. The welding strength of the welded portion 34 was 13.9 N / 15 mm.
[0097]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (T-180T manufactured by NBC Meshtec, pore diameter 86 μm, fiber diameter 55 μm, pore size 37%, Tm = 255 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to form a filter 16 for filtration. Obtained. The welding strength of the welded portion 34 was 21.6 N / 15 mm.
[0098]
Made of PET between two
　low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm). A filter 14 (T-100T manufactured by NBC Meshtec, pore diameter 183 μm, fiber diameter 71 μm, opening rate 52%, Tm = 255 ° C., length 80 mm, width 15 mm) is sandwiched and welded in this state to form a filter 16 for filtration. Obtained. The welding strength of the welded portion 34 was 32.1 N / 15 mm.
　The results of Examples 9 to 18 and Comparative Examples 8 to 18 are summarized in Table 3.
[0099]
[Table 3]

[0100]
　Hereinafter, in order to confirm the welding strength when a filter having a virtual opening rate of 100% is sandwiched, measurements were made with a sample in which two films were laminated and welded without sandwiching the filter 14.
[0101]
After stacking two
　high-density polyethylene films (HDPE, HD made by Tamapoli, Tm = 131 ° C., density 0.912 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) and welding on both sides. , The welding strength was measured. The welding strength was 51.8 N / 15 mm.
[0102]

　Two linear low-density polyethylene films (LLDPE, HC # 100 manufactured by Mitsui Chemicals Tohcello, Tm = 124 ° C., density 0.922 g / cm 3 , film thickness 100 μm, length 80 mm, width 15 mm) are laminated. To prepare a film having a film thickness of 200 μm. In this way, two linear low-density polyethylene films having a film thickness of 200 μm were laminated and welded on both sides, and then the welding strength was measured. The welding strength was 33.5 N / 15 mm.
[0103]

　Two low-density polyethylene films (LDPE, Tamapoli V-2, Tm = 112 ° C., density 0.923 g / cm 3 , film thickness 200 μm, length 80 mm, width 15 mm) are laminated and welded on both sides. After that, the welding strength was measured. The welding strength was 27.9 N / 15 mm.
　The results of Comparative Examples 19 to 21 are summarized in Table 4.
[0104]
[Table 4]

[0105]

　Regarding the relationship between the welding strength of the film material and the PET filter 14, as can be seen from Table 1, the film material using HDPE has the highest welding strength, and the one using LDPE has the highest welding strength. It can be seen that the lowest is the one using LLDPE and the welding strength during that period. The same thing can be seen from the results in Table 2 in which nylon was used as the material for the filter 14.
[0106]
　Regarding the effect of the film thickness on the welding strength between the film and the filter 14, as shown in Comparative Example 1-4 of Table 1, when LDPE is used as the film material, the film thickness has almost no effect. On the other hand, when LLDPE was used as the film material, it was found that the welding strength was higher when the film thickness was 200 μm than when it was 100 μm.
[0107]
　As described above, the fact that the film thickness dependence on the welding strength changes depending on the material of the filter 14 has not been elucidated in the mechanism of the phenomenon of welding used in the technique of the present disclosure, and the technique of the present disclosure is predictable. It indicates that it is related to a low-quality technical field.
[0108]
　From Table 3, it can be seen that the porosity of the filter 14 affects the welding strength. That is, when the opening rate of the filter 14 is in the range of about 10% to 40%, the higher the opening rate, the higher the welding strength with the film tends to be. However, when the film material is HDPE or LLDPE, the opening rate tends to be higher. There was also a tendency for the welding strength to saturate at a rate of about 40%. If the opening rate of the filter 14 exceeds 80%, the function as a filter cannot be established.
[0109]

D. Fabrication of container with filter A frame (size: outer frame 300 mm x 200 mm, inner frame) made of
　high-density polyethylene film (HDPE, manufactured by Hiroyuki Hosokawa, POLYELITE EH, Tm = 126 ° C, density 0.947 g / cm 3 , film thickness 200 μm) A PET filter 14 (Sefar 07-27 / 19, hole diameter 27 μm, hole opening rate 19%, Tm = 256 ° C.) is sandwiched between 270 mm × 170 mm, frame width is 15 mm at both ends), and the peripheral part is impulsed. Welding was performed at 230 ° C. using a sealer. As shown in FIG. 4, the filtration filter 16 having the PET filter thus produced is welded to the inner surface of the container 20 made of high-density polyethylene sheet by three sides, respectively, to form a PET filter. The filtration filter 16 having 14 was fixed between two high-density polyethylene sheets. Further, the polyethylene injection port 30 and the injection port 32 were attached to the high-density polyethylene sheet by welding, and a polyvinyl chloride tube 36 was attached to each port. Finally, the edge portions of the high-density polyethylene sheet other than the portion sandwiching the filtration filter 16 by the PET filter 14 are welded to each other, so that the filtration filter 16 by the PET filter 14 is inside the container 20. A container with a filter 22 welded to the above was prepared.
[0110]
　Compressed air of 0.01 MPa was introduced from the polyvinyl chloride tube 36 of the prepared container 22 with a filter through the injection port 30. At this time, the dispensing port 32 on the dispensing side was opened, and the bag internal pressure was adjusted to 0.01 MPa. By submerging the inflated container with a filter 22 into water, air leakage from the welded portion was confirmed, but no air leakage from the container with a filter 22 was observed.
[0111]
(Action / Effect of
　the present embodiment ) Next, the action and effect of the present embodiment will be described.
[0112]
　As described above, the filtration filter 16 includes a first welding frame 10, a second welding frame 12 arranged to face the first welding frame 10 in the thickness direction, and a first welding frame 10 having an outer peripheral portion thereof. It is composed of a filter 14 welded while being sandwiched between the entire circumference of the second welding frame 12 and the entire circumference of the second welding frame 12. Then, the first welding frame 10 is formed in a frame shape having a first passing hole 10A penetrated in the thickness direction with a flexible film having a film thickness of 120 μm or more made of a polymer, and the second welding frame 12 is formed. Is formed in a frame shape having a second passage hole 12A penetrated in the thickness direction with a flexible film having a film thickness of 120 μm or more made of a polymer. Further, the filter 14 is made of a material having a melting point higher than that of the first welding frame 10 and the second welding frame 12, having holes, and having a pore opening rate of 10% or more and 80% or less. , The melted first welding frame 10 and the second welding frame 12 from both sides of the filter 14 are connected through the holes 14A of the unmelted filter 14. Therefore, the first welding frame 10 and the second welding frame 12 facing each other can be bonded to each other to firmly fix the filter 14. As a result, since the filter 14, the first welding frame 10, and the second welding frame 12 are welded with high welding strength, the welding portion 34 is less likely to be peeled off, and the possibility of fluid leaking from the welding portion 34 is reduced.
[0113]
　Further, the first welding frame 10 and the second welding frame 12 are high-density polyethylene having a melting point of 120 ° C. to 140 ° C., linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., or a mixture of each. By being composed of the polymer containing the welded portion 34, the welded portion 34 is less likely to be peeled off.
[0114]
　Further, the first welding frame 10 and the second welding frame 12 are made of high-density polyethylene having a melting point of 120 ° C. to 140 ° C., linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., or a mixture of each. By being composed of the polymer, the welded portion 34 is more difficult to peel off.
[0115]
　Furthermore, the fluid in the filtration target can move in the order of the first passage hole 10A, the portion of the filter 14 that is not in contact with any of the first welding frame 10 and the second welding frame 12, and the second passage hole 12A. Since the first welding frame 10, the filter 14, and the second welding frame 12 are welded in the form, the fluid can be reliably flowed through the filter 14.
[0116]
　Further, the strength of the filter 14 itself can be ensured by configuring the filter 14 to contain at least one of polyester, polyamide, polyolefin, polyetheretherketone, polyethersulfone, carbon fiber, and metal.
[0117]
　Further, since the filter 14 is a woven fabric or a knitted fabric, the filter 14 has flexibility. Since the filter 16 is welded so as to partition the inside of the container 20 made of polymer, the container 22 with a filter can be easily shaken or kneaded, and the cell suspension can be obtained. Foreign matter contained in the turbid liquid can be easily removed.
[0118]
　Furthermore, since the pore size of the pore portion 14A in the filter 14 is 10 to 200 μm, it is possible to capture impurities and target cells while preventing clogging of the filter 14.
[0119]
　Further, since the filter 14 is formed in the shape of a rectangular sheet, the processing at the time of manufacturing the filter 14 becomes easy.
[0120]
　Further, since the shapes of the first welding frame 10 and the second welding frame 12 are substantially the same, the work efficiency at the time of manufacturing the filtration filter 16 can be improved.
[0121]
　Furthermore, the container 20 is formed by facing two rectangular polymer sheets having substantially the same shape and welding the peripheral edges of the polymer sheets to each other, and the filtering filter 16 provided inside the container 20. The first welding frame 10 is welded to one polymer sheet of the container 20 by a substantially U-shaped first welding portion 24 in which one is opened in the thickness direction, and the second welding in the filtering filter 16 is performed. The frame 12 is welded to the other polymer sheet of the container 20 by a substantially U-shaped second welded portion 26 in which the other is opened in the thickness direction, and the opening of the first welded portion 24 and the second welding The opening of the portion 26 is oriented in the opposite direction. Therefore, the fluid flowing from the opening of the first welding portion 24 to the filter 14 flows to the outside from the opening of the second welding portion 26 without significantly changing the direction of the flow. That is, since the fluid easily flows, impurities and target cells can be efficiently captured.
[0122]
　Further, by providing the container 20 with at least one of the injection port 30 and the ejection port 32, it is possible to easily put the fluid inside the container 20 and the inside of the container 20 partitioned by the filtration filter 16. By providing the injection port 30 on one side and the ejection port 32 on the other side, the cell suspension is injected into one compartment S1 inside the container 20 and the foreign matter is removed from the other compartment S2. It becomes easier to collect the turbid liquid. Further, the cell suspension is injected into one compartment S1 inside the container 20 in the filter-equipped container 22, the cells are filtered out by the filtration filter 16, and the liquid for resuspending the cells is injected into one compartment S1. It also facilitates resuspension of the cells and recovery of the cell suspension from one compartment S1.
[0123]
　Further, the filter 14 is sandwiched between the entire circumference of the first welding frame 10 and the entire circumference of the second welding frame 12, and is sandwiched between the first welding frame 10 and the second welding frame 12, respectively. The holes of the filter 14 in which the melted polymers of the first welding frame 10 and the second welding frame 12 are not melted because they are welded at a temperature equal to or higher than the melting point of 10 or the melting point of the second welding frame 12 and lower than the melting point of the filter 14. It can penetrate 14A and then solidify the polymer, for example by heat dissipation. As a result, the polymers of the first welding frame 10 and the second welding frame 12 that have penetrated into the hole 14A serve as anchors, so that the welding strength between the filter 14 and the first welding frame 10 and the second welding frame 12 is improved. be able to.
[0124]
　In the container with a filter 22 described above, the filter 14 is sandwiched between the first welding frame 10 and the second welding frame 12, but the present invention is not limited to this, and the container 20 is not shown. The filter 14 is directly welded to the inside of the container 20 so as to partition the container 20, and the space composed of one surface of the filter 14 and the inside of the container 20 is composed of the other surface of the filter 14 and the inside of the container 20. The inside of the container 20 may be partitioned by the filter 14 in such a manner that a flow path for moving the fluid to the space to be moved does not occur except for the filter 14. With such a configuration, it is possible to function as a filter while reducing the number of components.
Industrial applicability
[0125]
　The filter-equipped container 22 in which the filtration filter 16 of the technique of the present disclosure is incorporated in the container 20 is used, for example, for removing foreign substances in a cell suspension or washing cells. Therefore, the filter 16 for filtration and the container 22 with a filter using the filter 16 according to the technique of the present disclosure can be used, for example, in the manufacturing industry of filtration equipment.
[0126]
　The disclosure of Japanese Patent Application No. 2018-199182, filed October 23, 2018, is incorporated herein by reference in its entirety. All documents, patent applications, and technical standards described herein are to the same extent as if the individual documents, patent applications, and technical standards were specifically and individually stated to be incorporated by reference. Incorporated herein by reference.
The scope of the claims
[Claim 1]
　A first welding frame made of a flexible film containing a polymer and having a thickness of 120 μm or more and having a first passage hole penetrated in the thickness direction inside,
　and a thickness of 120 μm containing a polymer. A second passage hole composed of the above flexible film, arranged to face the first welding frame in the thickness direction, and penetrated in the thickness direction at a position corresponding to the first passage hole. It has a second welding frame formed in a frame shape on the inside, a
　melting point higher than that of the first welding frame and the second welding frame, and has holes, and the opening rate by the holes is 10% or more and 80% or less. The first welding frame and the second welding frame are made of the above-mentioned materials, and the outer peripheral portion is sandwiched between the entire circumference of the first welding frame and the entire circumference of the second welding frame. anda filter are welded respectively to the frame,
　said first welded frame, high density polyethylene, linear low density polyethylene having a melting point of 105 ° C. ~ 125 ° C. melting point of 120 ° C. ~ 140 ° C., or A
　filter for filtration, which is composed of a polymer containing a mixture of at least one of the high-density polyethylene and the linear low-density polyethylene .
[Claim 2]
　The first welding frame is a high-density polyethylene having a melting point of 120 ° C. to 140 ° C., a linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., or at least one of the high-density polyethylene and the linear low-density polyethylene.
　The filter for filtration according to claim 1, which is composed of a polymer composed of a mixture of each of the above.
[Claim 3]
　The fluid in the filtration target can move in the order of the first passage hole, the portion of the filter that is not in contact with any of the first welding frame and the second welding frame, and the second passage hole. The
　filtration filter according to claim 1 or 2, wherein the first welding frame, the filter, and the second welding frame are welded .
[Claim 4]
　The second welding frame is a high-density polyethylene having a melting point of 120 ° C. to 140 ° C., a linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., or at least one of the high-density polyethylene and the linear low-density polyethylene.
　The filter for filtration according to any one of claims 1 to 3 , which is composed of a polymer containing a mixture of the above.
[Claim 5]
　The second welding frame is a high-density polyethylene having a melting point of 120 ° C. to 140 ° C., a linear low-density polyethylene having a melting point of 105 ° C. to 125 ° C., or at least one of the high-density polyethylene and the linear low-density polyethylene.
　The filter for filtration according to any one of claims 1 to 3 , which is composed of a polymer composed of a mixture of the above.
[Claim 6]
　The filter for filtering according
　to any one of claims 1 to 5, wherein the filter contains at least one of polyester, polyamide, polyolefin, polyether ether ketone, polyether sulfone, carbon fiber, and metal . filter.
[Claim 7]
　The filter
　for filtration according to any one of claims 1 to 6, wherein the filter is a woven fabric or a knitted fabric .
[Claim 8]
　The filter
　for filtration according to any one of claims 1 to 7, wherein the pore diameter of the pore portion in the filter is 5 to 200 μm .
[Claim 9]
　The filter
　for filtration according to any one of claims 1 to 8, wherein the filter has a rectangular sheet shape .
[Claim 10]

　The filtration filter according to any one of claims 1 to 9, 　wherein the shapes of the first welding frame and the second welding frame are substantially the same .
[Claim 11]
　The filter is sandwiched between the entire circumference of the first welding frame and the entire circumference of the second welding frame, and is placed on the first welding frame and the second welding frame, respectively. The filtering filter according
　to any one of claims 1 to 10 , which is welded by heating at a temperature equal to or higher than the melting point of the second welding frame and lower than the melting point of the filter.
[Claim 12]
　A container with a filter, wherein the filtration filter according to any one of claims 1 to 11 is welded so as to partition the inside of the container made of a polymer
　.
[Claim 13]
　The container is formed by facing two rectangular flexible polymer sheets having substantially the same shape and welding the peripheral edges of the polymer sheets to each other, and is
　used in the filtration filter provided inside the container. The first welding frame is welded to one of the flexible polymer sheets of the container by a substantially U-shaped first welding portion in which one is opened in the thickness direction, and the first welding frame is formed in the
　filtering filter. The second welding frame is welded to the other flexible polymer sheet of the container by a substantially U-shaped second welding portion in which the other is opened in the thickness direction, and
　the first welding portion. 12. The
　container with a filter according to claim 12, wherein the opening of the second welded portion and the opening of the second welded portion are oriented in opposite directions .
[Claim 14]

　The filter-equipped container according to claim 12 or 13, 　wherein the container is provided with at least one of an injection port and an injection port .
[Claim 15]

　The filter-equipped container according to claim 14, further 　comprising an injection port on one side of the inside of the container partitioned by the filtration filter and an injection port on the other side .
[Claim 16]
　A container composed of two polymer sheets facing each other and welding peripheral portions of the polymer sheets to each other, and a container
　that is welded to the inside of the container so as to partition the inside of the container, and is provided with a hole. A
　container with a filter having a filter made of a material having a hole opening rate of 10% or more and 80% or less .
[Claim 17]
　A cell suspension is injected into one compartment inside the container in the container with a filter according to any one of claims 12 to 16, and the cell suspension is passed through a filter for filtration, and the other. A
　method of removing foreign matter contained in a cell suspension, which comprises the step of recovering a filtrate containing cells from the compartment .
[Claim 18]
　A cell suspension is injected into one compartment inside the container in the filter-equipped container according to any one of claims 12 to 16, and the cell suspension is passed through a filter for filtration and the other. The filtrate containing the cells is collected from the compartment, and a solution for cell suspension is injected into one of the compartments to resuspend the cells remaining in the compartment, and the resuspended cell suspension is used. A
　method for removing foreign substances contained in a cell suspension, which comprises a step of collecting a filtrate containing cells from the other compartment by passing the mixture through a filter for filtration .