Pipe sheet, storage container, cell generation system, processing container, and container

Abstract

A pipe sheet (1) has: a sheet body (10); a first pipe (21) that is provided inside the sheet body (10) and that is for performing a first step in a cell culturing method; a second pipe (22) that is provided inside the sheet body (10) and is for performing a second step in the cell culturing method; first joints (30) that are provided to one opening end of the first pipe (21) and one opening end of the second pipe (22) and are detachably linked to another member; and second joints (40) that are provided to the other opening end of the first pipe (21) and the other opening end of the second pipe (22), and are detachably linked to another member. A first joint (30) of the first pipe (21) and a first joint (30) of the second pipe (22) are positioned at a first end (10a) of the sheet body (10). A second joint (40) of the first pipe (21) and a second joint (40) of the second pipe (22) are positioned at a second end (10b) of the sheet body (10).

Description

Piping sheet, storage container, cell production system, processing container, and container

[0001] The present disclosure relates to a piping sheet used when performing a cell culture method or a cell production method, a storage container for storing cells cultured using the piping sheet, a cell production system for performing a cell production method, and a processing container used when performing a cell production method.

[0002] The present disclosure also relates to a container, and in particular to a container for processing cells or a closed container for processing cells.

[0003] Human stem cells, such as iPS cells (induced pluripotent stem cells) or ES cells (embryonic stem cells), are known as pluripotent cells that can be produced from tissue cells contained in human skin, organs, blood, etc. In particular, iPS cells can be produced using cells derived from the patient to be treated and then differentiated into cells of various tissues, and therefore are expected to be a transplant material for autologous transplants in regenerative medicine, which have a low rejection rate.

[0004] For example, when producing iPS cells from blood, hematopoietic stem cells are extracted from the blood and then infected with a viral vector. This allows iPS cells to be produced by introducing an iPS cell-initiating gene into the hematopoietic stem cells. Furthermore, when using the iPS cells obtained in this manner as transplantation materials, the iPS cells are cultured and expanded. The expanded iPS cells can then be induced to differentiate into T cells, which can then be used as immune cells, such as individual anti-cancer T cells.

[0005] When growing iPS cells created from hematopoietic stem cells, studies are being conducted on the use of cell culture devices to automatically culture and grow iPS cells. In this case, iPS cells can be grown by supplying medium to a culture vessel containing the iPS cells.

[0006] There are two main types of cell culture devices (automated culture devices): open-type cell culture devices and closed-type cell culture devices. In the open-type cell culture device, an open culture vessel such as a container or plate with an openable lid is used when culturing target cells (e.g., iPS cells). On the other hand, in the closed-type cell culture device, a closed culture vessel and closed piping are used when culturing target cells (Patent Document 1).

[0007] JP 2012-217435 A International Publication No. 2017 / 169259 JP 10-179137 A JP 2018-19630 A

[0008] However, in a sealed cell culture device, the piping (flow paths) must be replaced every time a process is performed, which makes the process of culturing or generating cells complicated and can lead to contamination.

[0009] The present disclosure is intended to solve such problems, and its first objective is to provide a piping sheet or the like that makes it easy to perform work when culturing or producing cells and that can prevent the occurrence of contamination.

[0010] Furthermore, when culturing cells such as iPS cells, as shown in (a) of Fig. 34, cells 2002 and culture medium 2003 are placed in a container 2001X such as a petri dish. Then, by periodically replacing or adding culture medium 2003, cells 2002 are proliferated as shown in (b) of Fig. 34. Once cells 2002 have proliferated, they are passaged as shown in (c) of Fig. 34. In other words, the proliferated cells 2002 are divided and transferred to multiple containers 2001X.

[0011] However, passaging cells is not only time-consuming but also has the risk of contamination during the passaging process. Therefore, a technique for culturing cells using a container with a variable volume has been proposed (e.g., Patent Document 2).

[0012] However, simply changing the volume of the container when culturing cells does not allow for efficient cell growth. For example, when infecting cells and then moving on to the culturing process, a small number of cells are cultured in suspension for infection, and then a large number of cells are cultured in adherent culture for growth. For this reason, it is desirable to increase the bottom surface area of ​​the container. Furthermore, changing the volume of the container at an inconsequential time does not allow for efficient cell growth.

[0013] The present disclosure also solves these problems, and has a second object to provide a container that can efficiently process cells.

[0014] Furthermore, when culturing cells such as iPS cells, the medium and the cells are housed in a container, and it has been known to use a closed container as a container for culturing cells in order to prevent contamination during cell culture (Patent Documents 3 and 4).

[0015] When processing cells, contents such as blood and cells are stored in a container, so there is a risk that the used container after cell processing will be contaminated with the contents, and there is a risk of infection when disposing of the used container. Therefore, there are restrictions on how used containers can be disposed of. For example, when disposing of a used container, it is necessary for the user to avoid touching any part of the container that has come into contact with contaminated materials.

[0016] Furthermore, to prevent contamination, cell processing containers are used up in one processing session, but if the used containers themselves are discarded, the amount of containers discarded will increase, which will have a negative impact on the environment.

[0017] The present disclosure also solves these problems, and its third objective is to provide a container that can be disposed of without touching any areas inside the container that have come into contact with contaminants, while also minimizing adverse effects on the environment.

[0018] In order to achieve the first object, one aspect of the piping sheet according to the present disclosure is a piping sheet used in carrying out a cell culture method including a first step and a second step, the piping sheet having a sheet-like sheet body, a first pipe provided inside the sheet body for carrying out the first step, a second pipe provided inside the sheet body for carrying out the second step, a first joint provided at one open end of the first pipe and one open end of the second pipe, each of which is detachably connected to another member, and a second joint provided at the other open end of the first pipe and the other open end of the second pipe, each of which is detachably connected to another member, wherein the first joint of the first pipe and the first joint of the second pipe are each located at a first end of the sheet body, and the second joint of the first pipe and the second pipe are each located at a second end of the sheet body.

[0019] In addition, in order to achieve the first object, one aspect of the storage container according to the present disclosure is a closed system storage container for storing cells cultured using the above-mentioned piping sheet, the storage container including a first space into which a liquid containing the cells flows, and a plurality of second spaces branched off from the first space.

[0020] Furthermore, in order to achieve the first object, one aspect of the cell generation system according to the present disclosure is a cell generation system for carrying out a cell generation method including a mutation step of generating mutated iPS cells by introducing an induction factor into a portion of cultured iPS cells to mutate them, and a differentiation induction step of mixing another portion of the cultured iPS cells with the mutated iPS cells to induce differentiation, the cell generation system comprising a single closed processing device for carrying out the mutation step and the differentiation induction step.

[0021] Furthermore, in order to achieve the first object, one aspect of the processing container according to the present disclosure is a processing container used when generating mutant iPS cells by introducing an induction factor into cultured iPS cells, and comprises a sealed container body and a plurality of cell compartments installed inside the container body, the container body being configured so that the iPS cells set in the plurality of cell compartments can be observed from the outside, the container body being configured so that an induction factor introducer that introduces induction factors into the plurality of cell compartments can access the inside of the container body, and the container body being configured so that a cell extractor that extracts iPS cells from the plurality of cell compartments can access the inside of the container body.

[0022] In order to achieve the second object, one aspect of the first container according to the present disclosure is a container for processing cells, the bottom of which deforms depending on the processing stage.

[0023] Furthermore, in order to achieve the third object, one aspect of the second container according to the present disclosure is a closed container for processing cells, which has a container body and a protective layer adhered to the inner surface of the container body, and the protective layer can be peeled off from the container body by physical or chemical methods.

[0024] The piping sheet, storage container, cell production system, and processing container according to the present disclosure make it easy to perform the work of culturing or producing cells and can prevent contamination from occurring.

[0025] Furthermore, according to the first container of the present disclosure, the bottom of the container can be deformed depending on the processing stage, allowing cells to be processed efficiently.

[0026] Furthermore, according to the second container of the present disclosure, the protective layer can be peeled off from the container body to remove the areas inside the container that have come into contact with contaminants, allowing the container to be disposed of without touching the areas inside the container that have come into contact with contaminants and reducing adverse effects on the environment.

[0027] FIG. 1 is a diagram showing a schematic configuration of a cell culture device according to embodiment 1. FIG. 2 is a perspective view of a piping sheet according to embodiment 1. FIG. 3A is a diagram for explaining a cell extraction step in the cell culture method according to embodiment 1. FIG. 3B is a diagram for explaining a first cell transfer step in the cell culture method according to embodiment 1. FIG. 3C is a diagram for explaining a gene transfer step in the cell culture method according to embodiment 1. FIG. 3D is a diagram for explaining a second cell transfer step in the cell culture method according to embodiment 1. FIG. 3E is a diagram for explaining a cell culture step in the cell culture method according to embodiment 1. FIG. 3F is a diagram for explaining a cell preservation step in the cell culture method according to embodiment 1. FIG. 4 is a diagram showing the configuration of a first modified storage container used in the cell culture device according to embodiment 1. FIG. 5 is a diagram showing the configuration of a second modified storage container used in the cell culture device according to embodiment 1. FIG. 6A is a side view of a storage container and a centrifuge in a modified cell culture device according to embodiment 1. FIG. 6B is a top view of a storage container in a modified cell culture device according to embodiment 1. FIG. 7 is a diagram showing a first example of an agitation mechanism used in the cell culture device according to embodiment 1. FIG. 8 is a diagram showing a second example of an agitation mechanism used in the cell culture device according to embodiment 1. FIG. 9 is a diagram showing a third example of an agitation mechanism used in the cell culture device according to embodiment 1. FIG. 10 is a diagram showing a fourth example of an agitation mechanism used in the cell culture device according to embodiment 1. FIG. 11 is a diagram for explaining the flow of a cell production method according to a modified example of embodiment 1. FIG. 12 is a diagram showing a schematic configuration of a cell production device according to a modified example of embodiment 1. FIG. 13 is a diagram for explaining a mutation step in the cell production method according to a modified example of embodiment 1. FIG. 14 is a perspective view of a container according to embodiment 2. FIG. 15 is a diagram showing how the container according to embodiment 2 is deformed. FIG. 16 is a diagram for explaining a container according to modified example 1 of embodiment 2. FIG. 17 is a diagram for explaining a container according to modified example 2 of embodiment 2. FIG. 18 is a diagram for explaining a container according to another aspect of modified example 2 of embodiment 2.FIG. 19 is a plan view of a scale used in a container according to another aspect of Modification 2 of Embodiment 2. FIG. 20 is a diagram for explaining a container according to Modification 3 of Embodiment 2. FIG. 21 is a diagram showing the configuration of a container according to Modification 4 of Embodiment 2. FIG. 22 is a perspective view of a container according to Embodiment 3. FIG. 23 is a cross-sectional view of a container according to Embodiment 3. FIG. 24 is a diagram for explaining a method of bonding a protective layer to a container body in a container according to Embodiment 3. FIG. 25 is a diagram for explaining a method of peeling a protective layer from a container body in a container according to Embodiment 3. FIG. 26 is a diagram for explaining a first modified example of a method of peeling a protective layer from a container body. FIG. 27 is a diagram for explaining a second modified example of a method of peeling a protective layer from a container body. FIG. 28 is a diagram showing an example of a method of discarding a peeled protective layer. FIG. 29 is a diagram showing the configuration of a container according to Modification 3 of Embodiment 3. FIG. 30 is a diagram for explaining a method of bonding a protective layer to a container body in a container according to Modification 3 of Embodiment 3. FIG. 31 is a diagram for explaining a method of peeling a protective layer from a container body in a container according to Modification 3 of Embodiment 3. Fig. 32 is a diagram showing the configuration of another aspect of a container according to a modification of embodiment 3. Fig. 33 is a diagram for explaining a method of peeling off the protective layer from the container body in another aspect of a container according to a modification of embodiment 3. Fig. 34 is a diagram for explaining a conventional cell culture method.

[0028] Specific embodiments of the present disclosure will be described below with reference to the drawings. Note that the embodiments described below all represent a comprehensive or specific example of the present disclosure. Therefore, the numerical values, shapes, materials, components, component placement and connection configurations, steps, and the order of steps shown in the following embodiments are merely examples and are not intended to limit the present disclosure. Therefore, among the components in the following embodiments, components not recited in independent claims will be described as optional components.

[0029] Furthermore, each drawing is a schematic diagram and is not necessarily a precise illustration. In each drawing, substantially the same components are denoted by the same reference numerals, and redundant explanations will be omitted or simplified.

[0030] First Embodiment First, a first embodiment of the present disclosure will be described with reference to Fig. 1 and Fig. 2. Fig. 1 is a diagram showing a schematic configuration of a cell culture device 100 according to the first embodiment. Fig. 2 is a perspective view of a piping sheet 1 according to the first embodiment.

[0031] The cell culture device 100 shown in Fig. 1 is a device for culturing target cells. The cell culture device 100 is a closed-type cell culture device in which various containers are connected by a piping sheet 1 so as to be constantly sealed.

[0032] In this embodiment, the target cells are iPS cells. Therefore, the cell culture device 100 has a cell culture mechanism for culturing and growing iPS cells. Furthermore, in this embodiment, iPS cells are produced from hematopoietic stem cells contained in blood. Specifically, iPS cells are produced by infecting hematopoietic stem cells extracted from blood with a viral vector and introducing an iPS cell conversion gene into the hematopoietic stem cells.

[0033] Furthermore, the cell culture device 100 not only has a cell culture mechanism for culturing iPS cells, but also has a cell extraction mechanism for extracting hematopoietic stem cells, which are the source of iPS cells, from blood, and a gene transfer mechanism for transferring iPS genes to the hematopoietic stem cells extracted from blood. In other words, by using the piping sheet 1, the cell culture device 100 can continuously perform a series of steps from extracting iPS cells from blood to culturing those iPS cells.

[0034] The target cells cultured by the cell culture device 100 are not limited to iPS cells. For example, T cells obtained by further inducing differentiation of iPS cells cultured by the cell culture device 100 may be used as target cells. In this case, the cell culture device 100 may have a mechanism capable of inducing differentiation of the cultured iPS cells. Furthermore, the target cells may be stem cells other than iPS cells, such as ES cells, or cells other than stem cells.

[0035] The cell culture device 100 has various containers, including a cell container 111, a bead container 112, a first culture medium container 113, a viral vector container 114, a second culture medium container 115, and a third culture medium container 116. The cell container 111, the bead container 112, the first culture medium container 113, the viral vector container 114, the second culture medium container 115, and the third culture medium container 116 are liquid containers that contain a predetermined liquid to be supplied to the piping sheet 1.

[0036] The cell culture device 100 also has various containers, including an extraction container 121, an infection container 122, a culture container 123, and a storage container 124. The extraction container 121, the infection container 122, the culture container 123, and the storage container 124 are liquid collection containers that collect liquid discharged from the piping sheet 1.

[0037] These liquid storage containers and liquid collection containers are closed bags and have a bag portion as the container body. The liquid storage containers and liquid collection containers are, for example, flexible transparent bags made of transparent resin film, but are not limited to this. For example, these liquid storage containers and liquid collection containers may be made of a resin material other than a transparent resin material, or may be made of a material other than a resin material, and may not be flexible. For example, the liquid storage containers and liquid collection containers may be containers made of glass or stainless steel.

[0038] Although not shown, the cell culture device 100 has a container mounting portion for mounting each of these containers. The container mounting portion has, for example, a structure that allows the container to be hooked and held. These containers are replaceable and can be attached to and detached from the container mounting portion. For example, these containers may be replaced each time target cells are cultured.

[0039] The cell container 111 is a container that contains a liquid that is a cell mixture containing first cells. In this embodiment, the cell container 111 contains blood that contains hematopoietic stem cells as the first cells.

[0040] The bead container 112 is a container that contains a liquid containing magnetic beads. Magnetic beads are an example of magnetic particles that adsorb to specific cells contained in a cell mixture. In this embodiment, the magnetic beads have the function of adsorbing to hematopoietic stem cells contained in blood.

[0041] The first culture medium container 113, the second culture medium container 115, and the third culture medium container 116 are containers that contain culture media for culturing cells. In this embodiment, since the target cells are iPS cells, the first culture medium container 113, the second culture medium container 115, and the third culture medium container 116 contain at least culture media for culturing iPS cells. The culture medium is a culture solution that contains nutrients and the like necessary for cell growth. The culture medium may be either a natural culture medium or a synthetic culture medium.

[0042] The viral vector container 114 is a container that contains a liquid containing a viral vector. A viral vector is a vector containing a virus that is used to transfer a specific gene to cells. In this embodiment, the viral vector is used to transfer an iPS cell transformation gene to hematopoietic stem cells contained in blood.

[0043] The extraction container 121 is a container for storing a liquid containing cells extracted from blood. The extraction container 121 is supplied with blood introduced from the cell container 111 into the piping sheet 1.

[0044] The infection container 122 is a container for infecting cells extracted from blood with a virus. The infection container 122 is supplied with cells extracted from blood and a liquid containing a viral vector.

[0045] The culture vessel 123 is a vessel for culturing cells. Cells are supplied to the culture vessel 123, and a culture medium for culturing the cells is also supplied. In other words, the culture vessel 123 contains cells and a culture medium. In this embodiment, the culture vessel 123 is a vessel for culturing iPS cells generated from hematopoietic stem cells contained in blood. Therefore, the culture vessel 123 contains at least hematopoietic stem cells, iPS cells, and a culture medium.

[0046] The storage container 124 is a container for collecting and storing target cells. In this embodiment, the target cells are iPS cells, and therefore the storage container 124 stores iPS cells that have been cultured and proliferated.

[0047] Next, a description will be given of the piping sheet 1. As shown in Figures 1 and 2, the piping sheet 1 includes a sheet-like sheet body 10, at least one pipe 20 provided inside the sheet body 10, and a first joint 30 and a second joint 40 connected to the pipe 20. The piping sheet 1 is a thin sheet with an integrated pipe, in which the sheet body 10 and the pipe 20 are integrated.

[0048] The piping sheet 1 is formed by bonding two sheets of sheet material. In this case, the sheet body 10 and the piping 20 are formed by the two sheets of sheet material, and the piping 20 is the spatial region between the two sheets of sheet material. In other words, the piping 20 (grooves) for the flow path, screws, etc. are processed in the two sheets of sheet material. A resin sheet made of a resin material can be used as the sheet material constituting the piping sheet 1. The sheet material may be a flexible sheet having flexibility, or a rigid sheet having no flexibility. The sheet material may also be transparent, translucent, or opaque. Note that the material constituting the sheet material is not limited to resin.

[0049] The piping sheet 1 is used when carrying out a cell culture method including a plurality of steps. In this embodiment, the cell culture method includes a cell extraction step (first step) of extracting cells, a gene imparting step (second step) of imparting genes to the cells, and a cell culture step (third step) of culturing the cells.

[0050] The sheet body 10 has a first end 10a and a second end 10b. The second end 10b is located opposite the first end 10a. In this embodiment, the sheet body 10 has a rectangular shape in plan view, and the first end 10a and the second end 10b are ends on a pair of long sides of the sheet body 10. Specifically, the first end 10a is an end on one side of the pair of long sides, and the second end 10b is an end on the other side of the pair of long sides. Note that the shape of the sheet body 10 in plan view is not limited to a rectangle.

[0051] The sheet body 10 has a pipe 20 inside. The portion of the sheet body 10 where the pipe 20 is not provided is flat. The thickness of the portion of the sheet body 10 where the pipe 20 is not provided is smaller than the diameter of the pipe 20.

[0052] The pipes 20 are flow paths through which a fluid flows. Specifically, a liquid or a gas flows through the pipes 20. The sheet main body 10 is provided with a plurality of independent pipes 20. Each of the plurality of pipes 20 may be straight or may be bent in an L-shape or the like. In other words, the shape of the pipes 20 that form the flow paths may be any shape.

[0053] In this embodiment, a first pipe 21, a second pipe 22, and a third pipe 23 are provided as the multiple pipes 20. The first pipe 21, the second pipe 22, and the third pipe 23 do not merge with each other and form independent flow paths.

[0054] The first pipe 21 is part of the cell extraction mechanism and is a pipe for performing the cell extraction step (first step). The first pipe 21 includes a first liquid pipe 21a through which mainly liquid flows and a first gas pipe 21b through which mainly gas flows.

[0055] The second pipe 22 is a part of the gene transfer mechanism and is a pipe for performing the gene transfer step (second step). The second pipe 22 includes a second liquid pipe 22a through which mainly liquid flows and a second gas pipe 22b through which mainly gas flows.

[0056] The third piping 23 is part of the cell culture mechanism and is a piping for performing the cell culture process (third process). The third piping 23 includes a third liquid piping 23a through which mainly liquid flows and a third gas piping 23b through which mainly gas flows.

[0057] It should be noted that not only liquid but also gas may flow through the first liquid pipe 21 a, the second liquid pipe 22 a, and the third liquid pipe 23 a. Also, not only gas but also liquid may flow through the first gas pipe 21 b, the second gas pipe 22 b, and the third gas pipe 23 b.

[0058] At least one of the first pipe 21, the second pipe 22, and the third pipe 23 includes a branch pipe having a branch path that branches off midway. The branch pipe is made up of a plurality of tributary pipes and a junction pipe where the plurality of tributary pipes join. In this embodiment, the first pipe 21 and the second pipe 22 include branch pipes. Specifically, the first liquid pipe 21a of the first pipe 21 and the second liquid pipe 22a of the second pipe 22 are branch pipes. The first liquid pipe 21a is made up of three tributary pipes and one junction pipe. The second liquid pipe 22a is made up of two tributary pipes and one junction pipe.

[0059] In this embodiment, the first gas pipe 21b of the first pipe 21, the second gas pipe 22b of the second pipe 22, and the third liquid pipe 23a and the third gas pipe 23b of the third pipe 23 are pipes that do not have branch paths.

[0060] Furthermore, the piping 20 includes connecting pipes 26 and 27 in addition to the first pipe 21, the second pipe 22, and the third pipe 23. The connecting pipe 26 connects the first pipe 21 and the second pipe 22. The connecting pipe 27 connects the second pipe 22 and the third pipe 23.

[0061] 1 , a plurality of first joints 30 and a plurality of second joints 40 are connected to the piping 20. Each of the plurality of first joints 30 is provided at a first end 10a of the seat main body 10. Each of the plurality of second joints 40 is provided at a second end 10b of the seat main body 10.

[0062] The multiple first joints 30 include first joints 31, 32, 33 and 34 provided at one open end of the first pipe 21 (the open end on the first end 10a side of the seat body 10), first joints 35, 36 and 37 provided at one open end of the second pipe 22 (the open end on the first end 10a side of the seat body 10), and first joints 38 and 39 provided at one open end of the third pipe 23 (the open end on the first end 10a side of the seat body 10).

[0063] Specifically, the open ends of the three branch pipes of the first liquid pipe 21a are located at the first end 10a of the seat body 10, and the first joints 31, 32, and 33 are provided at the open ends of the three branch pipes of the first liquid pipe 21a. Also, one open end of the first gas pipe 21b is located at the first end 10a of the seat body 10, and the first joint 34 is provided at one open end of the first gas pipe 21b.

[0064] The open ends of the two branch pipes of the second liquid pipe 22a are located at the first end 10a of the seat body 10, and the first joints 35 and 36 are provided at the open ends of the two branch pipes of the second liquid pipe 22a. Also, one open end of the second gas pipe 22b is located at the first end 10a of the seat body 10, and the first joint 37 is provided at one open end of the second gas pipe 22b.

[0065] One open end of the third liquid pipe 23a is located at the first end 10a of the seat body 10, and a first joint 38 is provided at one open end of the third liquid pipe 23a. Also, one open end of the third gas pipe 23b is located at the first end 10a of the seat body 10, and a first joint 39 is provided at one open end of the third gas pipe 23b.

[0066] The multiple second joints 40 include a second joint 41 provided at the other open end of the first pipe 21 (the open end on the second end 10b side of the seat body 10), a second joint 42 provided at the other open end of the second pipe 22 (the open end on the second end 10b side of the seat body 10), and a second joint 43 provided at the other open end of the third pipe 23 (the open end on the second end 10b side of the seat body 10).

[0067] Specifically, the open end of the junction pipe of the first liquid pipe 21a and the other open end of the first gas pipe 21b are located at the second end 10b of the seat body 10, and are both connected to the second joint 41. In this embodiment, the open end of the junction pipe of the first liquid pipe 21a and the other open end of the first gas pipe 21b join at the second joint 41 to form a single discharge pipe.

[0068] The open end of the junction pipe of the second liquid pipe 22a and the other open end of the second gas pipe 22b are located at the second end 10b of the seat body 10, and are both connected to the second joint 42. In this embodiment, the open end of the junction pipe of the second liquid pipe 22a and the other open end of the second gas pipe 22b join at the second joint 42 to form a single discharge pipe.

[0069] The other open end of the third liquid pipe 23a and the other open end of the third gas pipe 23b are located at the second end 10b of the seat body 10, and are both connected to the second joint 43. In this embodiment, the other open end of the third liquid pipe 23a and the other open end of the third gas pipe 23b join at the second joint 43 to form a single discharge pipe.

[0070] The second joints 40 include a second joint 44 provided midway on the connecting pipe 26 and a second joint 45 provided midway on the connecting pipe 27. The connecting pipe 26 is connected via the second joint 44 to a second joint 41 provided on the first pipe 21 and a second joint 42 provided on the second pipe 22. In this embodiment, the connecting pipe 26 connected to the second joint 41 and the connecting pipe 26 connected to the second joint 42 form independent discharge pipes at the second joint 44. The connecting pipe 27 is connected via the second joint 45 to a second joint 42 provided on the second pipe 22 and a second joint 43 provided on the third pipe 23. In this embodiment, the connecting pipe 27 connected to the second joint 42 and the connecting pipe 27 connected to the second joint 43 form independent discharge pipes at the second joint 45.

[0071] The first joint 30 and the second joint 40 are components that are detachably connected to another external member separate from the piping sheet 1. Specifically, the first joint 30 and the second joint 40 are one-touch joints, which allow the piping sheet 1 to be easily attached and detached from the external member. In this embodiment, the other member connected to the first joint 30 and the second joint 40 is any one of a liquid storage container, a liquid collection container, a pump, and a valve.

[0072] Specifically, in the first pipe 21, the first joint 31 is connected to the cell container 111 via a valve V1, the first joint 32 is connected to the bead container 112 via a valve V2, the first joint 33 is connected to the first culture medium container 113 via a valve V3, and the first joint 34 is connected to the pump P1. Also, in the first pipe 21, the second joint 41 is connected to the extraction container 121.

[0073] In the second piping 22, the first joint 35 is connected to the viral vector container 114 via a valve V4, the first joint 36 is connected to the second culture medium container 115 via a valve V5, and the first joint 37 is connected to a pump P2. In addition, in the second piping 22, the second joint 42 is connected to the infection container 122.

[0074] In the third piping 23, the first joint 38 is connected to the third culture medium container 116 via a valve V6, and the first joint 39 is connected to a pump P3. In addition, in the third piping 23, the second joint 43 is connected to the culture container 123.

[0075] The second joint 44 provided on the connecting pipe 26 is connected to a valve V7, and the second joint 45 provided on the connecting pipe 27 is connected to a valve V8.

[0076] Valves V1 to V8 are open / close valves. Valves V1 to V8 are, for example, pinch valves or air valves. By controlling valves V1 to V8, the flow of liquid or gas can be started or stopped. Pumps P1 to P3 control the liquid flowing through piping 20 of the piping sheet 1. Pumps P1 to P3 have the function of sucking up or sending liquid or gas. As an example, pumps P1 to P3 are decompression pumps. Valves V1 to V8 and pumps P1 to P3 are controlled by a controller included in the cell culture device 100. This allows cells to be cultured automatically according to a predetermined cell culture protocol without manual intervention.

[0077] Next, a cell culture method using the cell culture device 100 shown in Fig. 1 will be described with reference to Fig. 3A to Fig. 3F. Fig. 3A to Fig. 3F are diagrams for explaining the cell culture method according to embodiment 1. In each of the following steps, the open / closed states of valves V1 to V8 are appropriately controlled so that liquid or gas flows through a predetermined pipe 20.

[0078] 3A, piping sheet 1 is set in cell culture device 100, and liquid storage containers (cell container 111, bead container 112, first culture medium container 113, viral vector container 114, second culture medium container 115, third culture medium container 116) are connected to piping sheet 1. Furthermore, liquid collection containers (extraction container 121, infection container 122, culture container 123), which are empty containers, are connected to piping sheet 1. Furthermore, storage container 124 is connected to culture container 123.

[0079] Specifically, a cell container 111 containing blood including cells (hematopoietic stem cells) is connected to the first joint 31 of the piping sheet 1 via valve V1, a bead container 112 containing a liquid including magnetic beads is connected to the first joint 32 of the piping sheet 1 via valve V2, a first culture medium container 113 containing a culture medium is connected to the first joint 33 of the piping sheet 1 via valve V3, a viral vector container 114 containing a liquid including a viral vector is connected to the first joint 35 via valve V4, a second culture medium container 115 containing a culture medium is connected to the first joint 36 of the piping sheet 1 via valve V5, and a third culture medium container 116 containing a culture medium is connected to the first joint 38 of the piping sheet 1 via valve V6.

[0080] In addition, an extraction container 121, which is an empty container with no contents, is connected to the second joint 41 of the piping sheet 1, an infection container 122, which is an empty container with no contents, is connected to the second joint 42 of the piping sheet 1, and a culture container 123, which is an empty container with no contents, is connected to the second joint 43 of the piping sheet 1.

[0081] In addition, pumps P1, P2 and P3 are connected to the first joints 34, 37 and 39 of the piping sheet 1, and valves V7 and V8 are connected to the second joints 44 and 45 of the piping sheet 1.

[0082] In the cell culture method, first, cells contained in blood contained in the cell container 111 are extracted, and the extracted cells are contained in the extraction container 121 (cell extraction step).

[0083] Specifically, valve V1 is opened to supply blood contained in cell container 111 to piping sheet 1. The blood supplied to piping sheet 1 passes through first liquid pipe 21a of first pipe 21 and is supplied to extraction container 121 via second joint 41. Valve V2 is then opened to supply liquid containing magnetic beads contained in bead container 112 to piping sheet 1. The liquid containing magnetic beads supplied to piping sheet 1 passes through first liquid pipe 21a of first pipe 21 and is supplied to extraction container 121 via second joint 41. This causes the magnetic beads to adsorb to cells (hematopoietic stem cells) in extraction container 121. In other words, the cells in extraction container 121 are magnetized by the magnetic beads. Then, with valve V7 closed, pump P1 is operated to discharge the liquid from extraction container 121. This leaves only the cells in extraction container 121. In other words, the cells can be extracted into extraction container 121. Thereafter, valve V3 is opened to supply the culture medium contained in the first culture medium container 113 to the piping sheet 1. The culture medium supplied to the piping sheet 1 passes through the first liquid piping 21a of the first piping 21 and is supplied to the extraction container 121 via the second joint 41.

[0084] 3B, valve V7 is opened and pump P2 is operated to transfer the cells extracted in extraction container 121 together with the culture medium to empty infection container 122 (first cell transfer step). In this embodiment, the hematopoietic stem cells in extraction container 121 are transferred to empty infection container 122.

[0085] Next, as shown in FIG. 3C, the cells in the infection container 122 are infected by being imparted with the gene (gene imparting step).

[0086] Specifically, valve V4 is opened to supply a liquid containing a viral vector contained in viral vector container 114 to piping sheet 1. The liquid containing the viral vector supplied to piping sheet 1 passes through second liquid piping 22a of second piping 22 and is supplied to infection container 122 via second joint 42. This allows cells in infection container 122 to be infected with the virus and transformed into unique cells. In this embodiment, the cells in infection container 122 are hematopoietic stem cells, and therefore iPS cells are generated by infecting the hematopoietic stem cells with the virus.

[0087] If necessary, the culture medium may be supplied into the infection container 122 by opening the valve V5 before or after this step and supplying the culture medium contained in the second culture medium container 115 to the piping sheet 1.

[0088] 3D, valve V8 is opened and pump P3 is operated to move the unique cells generated in infection container 122 together with the culture medium to empty culture container 123 (second cell movement step). In this embodiment, iPS cells in infection container 122 are moved to empty culture container 123.

[0089] Next, as shown in FIG. 3E, the specific cells present in the culture vessel 123 are cultured (cell culture step).

[0090] Specifically, valve V6 is opened to supply the culture medium contained in the third culture medium container 116 to the piping sheet 1. The culture medium supplied to the piping sheet 1 passes through the third liquid piping 23a of the third piping 23 and is supplied to the culture vessel 123 via the second joint 43. This allows the unique cells in the culture vessel 123 to be cultured. In other words, the unique cells can be proliferated by cell division. In this embodiment, the unique cells in the culture vessel 123 are iPS cells, and therefore the iPS cells are cultured in the culture vessel 123.

[0091] It is preferable to supply the culture medium to the culture vessel 123 not all at once but in accordance with the cell division cycle of the specific cells. Moreover, since the medium changes over time due to metabolites and the like secreted by the specific cells, the old medium in the culture vessel 123 may be discharged and new medium may be supplied into the culture vessel 123 at an appropriate time during the culture period.

[0092] 3F, the cultured specific cells are then transferred from the culture vessel 123 to a storage vessel 124 (cell storage step). This allows the target cells to be stored in the storage vessel 124.

[0093] The used piping sheet 1 is discarded after the target cells are stored in the storage container 124. In other words, the piping sheet 1 is replaced every time target cells are cultured.

[0094] In this case, in the piping sheet 1 according to this embodiment, the first joint 30 is provided at the first end 10a of the sheet body 10, and the second joint 40 is provided at the second end 10b of the sheet body 10. Moreover, the first joint 30 and the second joint 40 are detachably connectable to other members. This configuration makes it easy to replace the piping sheet 1.

[0095] In particular, in the piping sheet 1 according to this embodiment, the first joint 30 and the second joint 40 are one-touch joints. This configuration makes it even easier to replace the piping sheet 1.

[0096] In the piping sheet 1 according to the present embodiment, piping 20 for carrying out a plurality of steps in the cell culture method is provided inside the sheet body 10. With this configuration, a plurality of steps in the cell culture method can be carried out consecutively using a single piping sheet 1. This makes it possible to easily carry out each step of the cell culture method and also to prevent contamination from occurring.

[0097] In the first embodiment, the closed storage container 124 for storing the cultured cells is a bag having one space (bag portion), but the present invention is not limited to this.

[0098] For example, as shown in FIG. 4, a closed storage container 200 for storing cells cultured on a piping sheet 1 may include a first space 210 into which a liquid containing cells flows, and a plurality of second spaces 220 branching off from the first space 210. The plurality of second spaces 220 are individually independent bag portions. In FIG. 4, five second spaces 220 are provided. The plurality of second spaces 220 have the same shape and can store the same amount of liquid. Furthermore, the five second spaces 220 are connected side by side, but are structured so that they can be easily separated by tearing.

[0099] By providing the plurality of second spaces 220 in the storage container 200, the same amount of cell-containing liquid can be divided and stored in a plurality of locations at approximately the same time while maintaining the closed state. For example, a plurality of cells with different uses, such as product cells, reference sample cells, and test sample cells, can be divided and stored in each of the plurality of second spaces 220.

[0100] Furthermore, since even a small difference in pressure loss in a microchannel tends to cause uneven flow in areas where the flow is easier, it is advisable to attach a pump P to each of the multiple second spaces 220, as shown in Figure 4. This allows the pump P to suck the liquid containing cells, making it possible to easily store the same amount of liquid in each of the multiple second spaces 220. In this case, instead of using multiple pumps P, the suction force may be adjusted for each second space 220, allowing one pump to suck the liquid containing cells into each of the multiple second spaces 220.

[0101] 4, by sealing across the plurality of second spaces 220, each of the plurality of second spaces 220 can be hermetically sealed. In other words, the storage container 200 can be divided into a non-storage area A1 and a storage area A2 with the sealing point S as the boundary.

[0102] 5, two adjacent second spaces 220 among the plurality of second spaces 220 may be spatially connected by a communication path 230. With this configuration, when different amounts of liquid flow from the first space 210 into the plurality of second spaces 220, the liquid that overflows from the second space 220 due to the inflow of more liquid is transferred to the adjacent second space 220 via the communication path 230. This makes it possible to store the same amount of liquid in the plurality of second spaces 220 at approximately the same time without using a pump. The communication path 230 is provided in the non-storage area A1.

[0103] 6A and 6B , by using the centrifuge 2 instead of a pump, liquid can be evenly introduced into each of the multiple second spaces 220. This allows the same amount of liquid to be stored in each of the multiple second spaces 220. FIG. 6A is a side view of the storage container 200B and the centrifuge 2. FIG. 6B is a top view of the storage container 200B. As shown in FIGS. 6A and 6B , the storage container 200B in this modification has a first space 210B into which the cell-containing liquid flows and multiple second spaces 220B branching off from the first space 210B. However, the multiple second spaces 220B are not connected to each other and are arranged separately in the rotation direction of the centrifuge 2.

[0104] Furthermore, when storing a liquid containing cultured cells in a storage container, the liquid containing the cells may be poured into the storage container while being stirred.

[0105] For example, as shown in FIG. 7 , in a storage container 200C having a first space 210C and a plurality of second spaces 220C, the liquid flowing into the storage container 200C may be agitated by a propeller 3 disposed in the first space 210C. Alternatively, as shown in FIG. 8 , the liquid flowing into the storage container 200C may be agitated by vibrating the storage container 200C using a vibration device such as an ultrasonic vibration device. Alternatively, as shown in FIG. 9 , the liquid flowing into the storage container 200C may be agitated by supplying bubbles to the first space 210C of the storage container 200C using an air bubble supply device. Note that instead of using an agitation device such as a propeller, vibration device, or air bubble supply device, the liquid flowing into the storage container 200C may be agitated by mixing the liquid using a mixing channel 4 (agitation channel) between the culture vessel 123 and the storage container 200C, as shown in FIG. 10 .

[0106] (Modification of First Embodiment) Next, a modification of the first embodiment will be described with reference to Figs. 11 to 13. Fig. 11 is a diagram illustrating the flow of a cell generation method according to a modification of the first embodiment. Fig. 12 is a diagram illustrating the schematic configuration of a cell generation device 100A according to a modification of the first embodiment. Fig. 13 is a diagram illustrating the mutation step in the cell generation method according to the modification of the first embodiment.

[0107] In this modification, cultured cells are mutated to generate a disease model, which allows the reproduction of the disease. Note that in this modification, iPS cells are used as the cells.

[0108] 11 , the cell generation method of this modification includes a culture step (a) of culturing iPS cells, a mutation step (b) of mutating some of the cultured iPS cells by introducing an inducer into them to generate mutant iPS cells, and a differentiation induction step (c) of mixing another part of the cultured iPS cells with the mutated iPS cells and inducing differentiation. In the mutation step (b), for example, a disease-associated gene mutation is introduced as an inducer, and genome editing is performed by electroporation.

[0109] The cell production device 100A shown in FIG. 12 is a cell production system for carrying out the cell production method shown in FIG.

[0110] The cell generation device 100A is a device for generating target cells. The cell generation device 100A is a closed-type cell generation device, with various containers connected by a piping sheet 1A so as to maintain a constant airtight state. The cell generation device 100A has a cell extraction mechanism, a gene transfer mechanism, a cell culture mechanism, a mutation mechanism, and a differentiation induction mechanism. The cell extraction mechanism, gene transfer mechanism, and cell culture mechanism are the same as those of the cell culture device 100 in embodiment 1. In other words, the cell generation device 100A in this modification has a structure in which a mutation mechanism and a differentiation induction mechanism are added to the cell culture device 100 in embodiment 1. Therefore, the process from extracting iPS cells from blood to culturing the extracted iPS cells is the same as embodiment 1. The cell generation device 100A can continuously perform a series of steps from inducing differentiation of iPS cells extracted from blood.

[0111] Similar to the first embodiment, the cell generation device 100A has a cell container 111, a bead container 112, a first culture medium container 113, a viral vector container 114, a second culture medium container 115, and a third culture medium container 116. The cell generation device 100A further has, as liquid containers, a fourth culture medium container 117, a differentiation induction factor container 118, and a fifth culture medium container 119. The fourth culture medium container 117 and the fifth culture medium container 119 are containers that contain culture media for culturing cells. The differentiation induction factor container 118 is a container that contains a liquid containing a differentiation induction factor.

[0112] Similarly to the first embodiment, the cell generation device 100A also includes an extraction container 121, an infection container 122, and a culture container 123. The cell generation device 100A further includes a disease mutation container 125, a differentiation induction container 126, and a storage container 127. The disease mutation container 125 is a closed system processing container for carrying out the mutation step (b) in the cell generation method. The differentiation induction container 126 is a closed system processing container for carrying out the differentiation induction step (c) in the cell generation method. The storage container 127 is the same as the storage container 124 in the first embodiment.

[0113] The cell generation device 100A includes a piping sheet 1A as a closed system processing device for performing at least the mutation step (b) and the differentiation induction step (c). In this modification, the piping sheet 1A can perform not only the mutation step and the differentiation induction step, but also the cell extraction step, gene transfer step, and cell culture step prior to the mutation step and the differentiation induction step.

[0114] The piping sheet 1A in this modified example, like the piping sheet 1 in embodiment 1 above, comprises a sheet-shaped sheet body 10, piping 20 provided inside the sheet body 10, and a first joint 30 and a second joint 40 connected to the piping 20.

[0115] In the piping sheet 1A of this modification, the piping 20 includes a fourth pipe 24 and a fifth pipe 25 in addition to the first pipe 21, the second pipe 22, and the third pipe 23. The fourth pipe 24 and the fifth pipe 25 do not merge with the first pipe 21, the second pipe 22, and the third pipe 23, and form independent flow paths.

[0116] The fourth pipe 24 is a part of the mutation mechanism and is a pipe for performing the mutation process. The fourth pipe 24 includes a fourth liquid pipe 24a through which mainly liquid flows and a fourth gas pipe 24b through which mainly gas flows.

[0117] The fifth pipe 25 is a part of the differentiation induction mechanism and is a pipe for performing the differentiation induction step. The fifth pipe 25 includes a fifth liquid pipe 25a through which mainly liquid flows and a fifth gas pipe 25b through which mainly gas flows.

[0118] It should be noted that not only liquid but also gas may flow through the fourth liquid pipe 24a and the fifth liquid pipe 25a, and not only gas but also liquid may flow through the fourth gas pipe 24b and the fifth gas pipe 25b.

[0119] The fourth pipe 24 and the fifth pipe 25 may include a branch pipe having a branch path that branches off midway. In this modification, the fifth pipe 25 includes a branch pipe. Specifically, the fifth liquid pipe 25a of the fifth pipe 25 is a branch pipe. The fifth liquid pipe 25a is composed of two branch pipes and one junction pipe. Note that the fourth liquid pipe 24a and the fourth gas pipe 24b of the fourth pipe 24 and the fifth gas pipe 25b of the fifth pipe 25 are pipes that do not have a branch path.

[0120] In this modification, the piping 20 includes not only the connecting piping 26 and 27 but also the connecting piping 28 and 29. The connecting piping 28 connects the third piping 23 and the fourth piping 24. The connecting piping 29 connects the fourth piping 24 and the fifth piping 25.

[0121] In addition to first joints 31 to 39, the multiple first joints 30 also include first joints 30a and 30b provided at one open end of the fourth pipe 24 (the open end on the first end 10a side of the seat body 10), and first joints 30c, 30d, and 30e provided at one open end of the fifth pipe 25 (the open end on the first end 10a side of the seat body 10).

[0122] Specifically, one open end of the fourth liquid pipe 24a is located at the first end 10a of the sheet body 10, and the first joint 30a is provided at one open end of the fourth liquid pipe 24a. Also, one open end of the fourth gas pipe 24b is located at the first end 10a of the sheet body 10, and the first joint 30b is provided at one open end of the fourth gas pipe 24b. The first joint 30a is connected to the fourth culture medium container 117 via a valve V9, and the first joint 30b is connected to a pump P4.

[0123] The open ends of the two branch pipes of the fifth liquid pipe 25a are located at the first end 10a of the sheet body 10, and first joints 30c and 30d are provided at the open ends of the two branch pipes of the fifth liquid pipe 25a. One open end of the fifth gas pipe 25b is located at the first end 10a of the sheet body 10, and first joint 30e is provided at one open end of the fifth gas pipe 25b. The first joint 30c is connected to a differentiation-inducing factor container 118 via a valve V10, the first joint 30d is connected to a fifth culture medium container 119 via a valve V11, and the first joint 30e is connected to a pump P5.

[0124] In addition to second joints 41 to 45, the multiple second joints 40 also include a second joint 40a provided at the other open end of the fourth pipe 24 (the open end on the second end 10b side of the seat body 10) and a second joint 40b provided at the other open end of the fifth pipe 25 (the open end on the second end 10b side of the seat body 10).

[0125] Specifically, the open end of the junction pipe of the fourth liquid pipe 24a and the other open end of the fourth gas pipe 24b are located at the second end 10b of the sheet body 10 and are both connected to the second joint 40a. The open end of the junction pipe of the fourth liquid pipe 24a and the other open end of the fourth gas pipe 24b join at the second joint 40a to form a single discharge pipe. The second joint 40a is connected to the disease mutation container 125.

[0126] The open end of the junction pipe of the fifth liquid pipe 25a and the other open end of the fifth gas pipe 25b are located at the second end 10b of the sheet body 10 and are both connected to the second joint 40b. The open end of the junction pipe of the fifth liquid pipe 25a and the other open end of the fifth gas pipe 25b join at the second joint 40b to form a single discharge pipe. The second joint 40b is connected to the differentiation induction container 126.

[0127] The second joints 40 include a second joint 40c provided midway on the connecting pipe 28 and a second joint 40d provided midway on the connecting pipe 29. The connecting pipe 28 is connected via the second joint 40c to a second joint 43 provided on the third pipe 23 and a second joint 40a provided on the fourth pipe 24. The connecting pipe 29 is connected via the second joint 40d to a second joint 40a provided on the fourth pipe 24 and a second joint 40b provided on the fifth pipe 25. The second joint 40c provided on the connecting pipe 28 is connected to the valve V12, and the second joint 40d provided on the connecting pipe 29 is connected to the valve V13.

[0128] Valves V9 to V13 are on-off valves, similar to valves V1 to V8. Pumps P4 and P5 are pressure reducing pumps, similar to pumps P1 to P3, and control the liquid flowing through the piping 20 of the piping sheet 1A. Valves V9 to V13 and pumps P4 and P5 are controlled by a controller included in the cell generation device 100A, similar to valves V1 to V8 and pumps P1 to P3. This allows target cells to be generated automatically according to a predetermined cell generation protocol without manual intervention.

[0129] Next, a cell generation method using the cell generation device 100A shown in FIG. 12 will be described.

[0130] The cell extraction step, gene transfer step, and cell culture step in the cell generation method according to this modification can first be performed according to the same flow as in the above-described embodiment 1. Specifically, iPS cells cultured in the culture vessel 123 can be obtained according to the flow shown in Figures 3A to 3E.

[0131] Next, valve V12 is opened and pump P4 is operated to move some of the iPS cells in culture container 123 together with the medium to disease mutation container 125. For example, 20% of the cultured iPS cells are moved to disease mutation container 125.

[0132] Next, an inducer is introduced into the iPS cells transferred to the disease mutation container 125 to mutate the iPS cells and generate mutant iPS cells (mutation step). Specifically, the disease mutation container 125 shown in Figure 13 is used to mutate the iPS cells and generate mutant iPS cells as disease cells.

[0133] 13, the disease mutation container 125 includes a sealed container body 125a and a plurality of cell compartments 125b disposed inside the container body 125a. The iPS cells placed in the plurality of cell compartments 125b are the iPS cells in the culture container 123 and have been cultured using the piping sheet 1A.

[0134] The container body 125a is configured so that the iPS cells set in the multiple cell compartments 125b can be observed from the outside. For example, a transparent observation window is provided in part of the container body 125a so that the inside of the container body 125a can be observed.

[0135] The container body 125a is configured so that an inducer introducer 310, which introduces inducers into the multiple cell compartments 125b, can access the inside of the container body 125a. In this modification, a through-hole 125c is provided in the container body 125a as an access hole through which the inducer introducer 310 can be inserted. The inducer introducer 310 is, for example, a pipette that can be operated by a drive unit.

[0136] Furthermore, the container body 125a is configured to allow a cell extractor 320, which extracts iPS cells from the multiple cell compartments 125b, to access the inside of the container body 125a. In this modification, a through-hole 125d is provided in the container body 125a as an access hole through which the cell extractor 320 can be inserted. The cell extractor 320 is, for example, a pipette that can be operated by a drive unit.

[0137] When mutating iPS cells using the disease mutation container 125 configured in this manner, an inducer introducer 310 is inserted into the container body 125a through the through-hole 125c, and an inducer is introduced into the iPS cells that have been moved to the multiple cell compartments 125b. Next, the iPS cells are genetically modified by electroporation to mutate the disease, resulting in mutant iPS cells. Next, the mutant iPS cells are identified using an observation device 400, such as a camera. Next, a cell extractor 320 is inserted into the container body 125a through the through-hole 125d, and the mutant iPS cells identified in the multiple cell compartments 125b are extracted by aspirating them with the cell extractor 320. Unnecessary iPS cells (iPS cells that have not been mutated) are then washed away from the container body 125a.

[0138] In the mutation process, if necessary, the culture medium may be supplied into the disease mutation container 125 by opening the valve V9 and supplying the culture medium contained in the fourth culture medium container 117 to the piping sheet 1A.

[0139] Next, valves V12 and V13 are opened and pump P5 is operated to move another portion of the iPS cells in the culture vessel 123 together with the culture medium to the differentiation induction vessel 126. In other words, the remaining iPS cells in the culture vessel 123 that were not moved to the disease mutation vessel 125 are moved to the differentiation induction vessel 126. For example, in this modification, since 20% of the iPS cells are moved to the disease mutation vessel 125 to generate mutant iPS cells, the remaining 80% of the iPS cells are moved to the differentiation induction vessel 126.

[0140] Next, the mutated iPS cells generated in the mutation step are mixed with the cultured iPS cells and induced to differentiate (differentiation induction step). Specifically, the iPS cells transferred from the culture container 123 to the differentiation induction container 126 are mixed with the mutated iPS cells extracted by the cell extractor 320 in the differentiation induction container 126, and valve V10 is opened to supply a liquid containing a differentiation induction factor contained in the differentiation induction factor container 118 to the piping sheet 1A. The liquid containing the differentiation induction factor supplied to the piping sheet 1A passes through the fifth liquid pipe 25a of the fifth pipe 25 and is supplied to the differentiation induction container 126 via the second joint 40b. This induces differentiation of the mutated iPS cells, which are disease-mutated cells.

[0141] At this time, if necessary, the valve V11 may be opened to supply the culture medium contained in the fifth culture medium container 119 to the piping sheet 1A, thereby supplying the culture medium into the differentiation induction container 126.

[0142] Next, the differentiation-induced mutant iPS cells (target cells) are transferred from differentiation induction container 126 to storage container 127. This allows the target cells to be stored in storage container 127. Transfer of mutant iPS cells from differentiation induction container 126 to storage container 127 can be performed in the same manner as the transfer of iPS cells from culture container 123 to storage container 124 in the first embodiment.

[0143] The used piping sheet 1A is discarded after the target cells are stored in the storage container 127. That is, in this modified example as well, the piping sheet 1A is replaced every time target cells are cultured.

[0144] In this case, the piping sheet 1A according to this modification also has a first joint 30 provided at the first end 10a of the sheet body 10 and a second joint 40 provided at the second end 10b of the sheet body 10. Furthermore, the first joint 30 and the second joint 40 are detachably connectable to other members. This configuration makes it easy to replace the piping sheet 1A.

[0145] Moreover, in the piping sheet 1A according to this modified example, the first joint 30 and the second joint 40 are also one-touch joints, which makes it even easier to replace the piping sheet 1A.

[0146] The piping sheet 1A according to this modification also has piping 20 for carrying out multiple steps in the cell production method provided inside the sheet body 10. This configuration allows multiple steps in the cell production method to be carried out consecutively using a single piping sheet 1A. This makes it easy to perform each step of the cell production method and also prevents contamination from occurring.

[0147] Furthermore, the cell generation device 100A according to this modification can realize a cell generation system suitable for disease models. In particular, by using the disease mutation container 125 as a closed processing container for performing the mutation step in the cell generation method, a cell generation system suitable for disease models can be realized.

[0148] (Embodiment 2) Next, the configuration of a container 1001 according to embodiment 2 will be described with reference to Fig. 14. Fig. 14 is a perspective view of the container 1001 according to embodiment 2.

[0149] The container 1001 shown in FIG. 14 is a container for processing cells. In this embodiment, the container 1001 is a cell culture container for culturing cells. For example, the container 1001 processes human-derived cells such as iPS cells. As an example, the container 1001 cultures human stem cells. By culturing the cells in the container 1001, the cells proliferate. Note that iPS cells are produced from hematopoietic stem cells contained in blood. Specifically, iPS cells can be obtained by infecting hematopoietic stem cells extracted from blood with a viral vector and introducing an iPS cell transformation gene into the virally infected hematopoietic stem cells.

[0150] As shown in FIG. 14, the container 1001 is a cylindrical container with a bottom having an opening 1001 a , and has a bottom 1010 having a bottom surface 1010 a and a side wall 1020 standing on the bottom 1010 .

[0151] The side wall portion 1020 is a low, flat, cylindrical body. The side wall portion 1020 is formed along the outer edge of the bottom surface 1010a of the bottom portion 1010, surrounding the bottom surface 1010a. The side wall portion 1020 is erected on the bottom portion 1010 in an orientation perpendicular to the bottom surface 1010a of the bottom portion 1010. The bottom portion 1010 and the side wall portion 1020 are formed in a flat plate shape with a constant thickness.

[0152] The shape of the bottom surface 1010a of the bottom portion 1010 in a plan view is polygonal. In this case, the side wall portion 1020 is a square tube. In this embodiment, the shape of the bottom surface 1010a of the bottom portion 1010 in a plan view is hexagonal. Therefore, the shape of the side wall portion 1020 in a top view is hexagonal. In other words, the opening shape of the opening 1001a of the container 1001 is hexagonal.

[0153] The bottom 1010 is configured to be deformable. Specifically, the bottom 1010 is made of an elastic material and can expand and contract. As an example, the bottom 1010 is an elastic member having rubber elasticity. For example, the bottom 1010 is made of an elastomer such as silicone rubber. It is preferable that the bottom 1010 has flexibility.

[0154] In this embodiment, the size of the bottom 1010 changes while remaining flat. Specifically, the bottom 1010 expands and contracts in a flat shape. When the bottom 1010 expands in a flat shape, the area of ​​the bottom surface 1010a of the bottom 1010 increases. When the bottom 1010 contracts in a flat shape, the area of ​​the bottom surface 1010a of the bottom 1010 decreases.

[0155] The side wall portion 1020 has a first side wall portion 1021 which is an expandable and contractible portion, and a second side wall portion 1022 which is less expandable than the first side wall portion 1021 .

[0156] The first side wall portion 1021 is made of a stretchable material and can expand and contract. As an example, the first side wall portion 1021 is an elastic member having rubber elasticity. For example, the first side wall portion 1021 is made of an elastomer such as silicone rubber. The first side wall portion 1021 may be flexible. In this embodiment, the first side wall portion 1021 is made of the same material as the bottom portion 1010, but the first side wall portion 1021 and the bottom portion 1010 may be made of different materials.

[0157] The second side wall portion 1022 is made of a material that is difficult to stretch. In this embodiment, the second side wall portion 1022 is basically a non-stretchable portion that does not stretch. The second side wall portion 1022 is made of, for example, a hard resin material or a metal material.

[0158] The first side wall portions 1021 and the second side wall portions 1022 are alternately provided in the circumferential direction along the outer edge of the bottom surface 1010a of the bottom portion 1010. Specifically, each of the first side wall portions 1021 and the second side wall portions 1022 corresponds to one side of the bottom surface 1010a. In this embodiment, since the bottom surface 1010a has a hexagonal shape in a plan view, the side wall portion 1020 is composed of three first side wall portions 1021 and three second side wall portions 1022. In other words, the bottom portion 1010 is surrounded by the three first side wall portions 1021 and three second side wall portions 1022. The first side wall portions 1021 and the second side wall portions 1022 each have a rectangular plate shape.

[0159] The volume of the container 1001 configured in this manner changes as the bottom 1010 deforms. In this embodiment, the container 1001 deforms at the bottom 1010 and the first side wall 1021. In other words, the container 1001 is a variable container in which the shapes of the bottom 1010 and the first side wall 1021 are variable.

[0160] Here, the characteristics of the container 1001 will be described with reference to Fig. 15. Fig. 15 is a diagram showing how the container 1001 according to the second embodiment is deformed.

[0161] 15 , a container 1001 is used when processing cells 1002. In this embodiment, the container 1001 is a culture container for culturing the cells 1002, and the container 1001 is used when culturing the cells 1002.

[0162] 15 , when cells 1002 are cultured, cells 1002 are supplied to a container 1001, along with a culture medium 1003 for culturing the cells 1002. As an example, the cells 1002 are iPS cells generated from hematopoietic stem cells contained in blood. In this case, the container 1001 contains the cells 1002, which are iPS cells, and the culture medium 1003.

[0163] The bottom 1010 of the container 1001 is deformed depending on the processing stage for processing the cells 1002. In this embodiment, the bottom 1010 of the container 1001 is deformed depending on the culture stage for culturing the cells 1002. That is, the timing when the bottom 1010 is deformed is the culture stage. For example, the culture stage at which the bottom 1010 is deformed is the transition from initial culture ( FIG. 15( a) ) to expansion culture ( FIG. 15( b) ). In this case, the bottom 1010 of the container 1001 is deformed when transitioning from initial culture to expansion culture. Note that in this embodiment, as shown in FIGS. 15( b) and 15( c ), the bottom 1010 of the container 1001 is further deformed during expansion culture. That is, the bottom 1010 of the container 1001 is also deformed when transitioning from first expansion culture ( FIG. 15( b) ) to second expansion culture ( FIG. 15( c) ).

[0164] Furthermore, the area of ​​the bottom surface 1010a of the bottom 1010 of the container 1001 changes depending on the culture stage. That is, the area of ​​the bottom surface 1010a of the bottom 1010 of the container 1001 changes as the bottom 1010 of the container 1001 deforms. Specifically, as shown in Fig. 15 , the area of ​​the bottom surface 1010a of the bottom 1010 of the container 1001 increases as the bottom 1010 expands and deforms into a planar shape depending on the culture stage. That is, the area of ​​the bottom surface 1010a of the bottom 1010 increases as the bottom surface 1010a of the bottom 1010 expands.

[0165] Furthermore, the bottom 1010 of the container 1001 is deformed by the deformation of the side wall 1020. That is, the bottom 1010 of the container 1001 is deformed in conjunction with the deformation of the side wall 1020. Specifically, the bottom 1010 is deformed by moving the second side wall 1022 of the side wall 1020. In this embodiment, moving the second side wall 1022 not only deforms the bottom 1010 but also deforms the first side wall 1021. That is, the bottom 1010 and the first side wall 1021 are deformed in conjunction with the movement of the second side wall 1022.

[0166] For example, as shown by the arrows in Figure 15, by pulling all of the second side walls 1022 outward, the second side walls 1022 are translated outward, and the first side walls 1021 and the bottom 1010 are stretched and deformed. In this case, the first side walls 1021 are stretched and deformed in their longitudinal direction, and the bottom 1010 is stretched and deformed in a planar manner. As the bottom 1010 is expanded and deformed in a planar manner, the area of ​​the bottom surface 1010a of the bottom 1010 increases. Note that even if the first side walls 1021 and the bottom 1010 are expanded and deformed, the second side walls 1022 do not deform.

[0167] The second side wall 1022 may be moved manually, but is preferably moved mechanically. For example, the first side wall 1021 and the bottom 1010 may be expanded by manually moving the second side wall 1022 outward by a user, or the first side wall 1021 and the bottom 1010 may be automatically expanded by moving the second side wall 1022 outward using a driving device or the like. In this way, by expanding the first side wall 1021 and the bottom 1010 using mechanical force, the bottom 1010 of the container 1001 can be easily deformed at a predetermined timing for a culture stage.

[0168] In this way, the container 1001 has the bottom 1010 and the first side wall 1021 expand and deform according to the culture stage, so that the volume can be expanded according to the culture stage.

[0169] The magnification of the bottom 1010 of the container 1001 is not particularly limited. As an example, if the area of ​​the bottom surface 1010a of the bottom 1010 is 1 when the area of ​​the bottom surface 1010a of the bottom 1010 is at its smallest (the state shown in FIG. 15(a)), the magnification of the bottom surface 1010a of the bottom 1010 is greater than 1 and equal to or less than 20. From the viewpoint of culturing cells, the magnification of the bottom surface 1010a of the bottom 1010 may be 5 to 10 times.

[0170] As described above, the container 1001 according to this embodiment is a container for processing the cells 1002, and the bottom 1010 of the container 1001 is deformed depending on the processing stage.

[0171] In this way, bottom 1010 of container 1001 can be deformed according to the processing stage rather than being deformed at meaningless timing, thereby enabling efficient processing of cells 1002. Specifically, in this embodiment, bottom 1010 of container 1001 is deformed according to the culture stage, allowing cells 1002 to be cultured and proliferated efficiently.

[0172] Furthermore, in this embodiment, the deformation of bottom 1010 of container 1001 changes the area of ​​bottom surface 1010a of bottom 1010 in container 1001. Specifically, the expansion and deformation of bottom 1010 of container 1001 increases the area of ​​bottom surface 1010a of bottom 1010.

[0173] In this way, by increasing the area of ​​the bottom surface 1010a of the bottom portion 1010, it is possible to efficiently carry out adhesion culture of the cells 1002. This allows the cells 1002 to be efficiently cultured and proliferated.

[0174] In this embodiment, the bottom 1010 of the container 1001 is deformed into a flat shape, but this is not limiting. For example, the bottom 1010 may be deformed into a dome shape that bulges downward.

[0175] In addition, in the present embodiment, the shape of the bottom 1010 of the container 1001 in a plan view is hexagonal, but it may be a polygon other than a hexagon. Furthermore, the shape of the bottom 1010 of the container 1001 in a plan view is not limited to a polygon and may be circular. In this case, the side wall 1020 is cylindrical, and first side wall portions 1021 and second side wall portions 1022 that are partially arc-shaped are alternately arranged along the circumferential direction.

[0176] Furthermore, in this embodiment, the container 1001 is a cylindrical container with a bottom and an opening 1001a, but this is not limiting. For example, the container 1001 may be a closed-type container having an upper surface (top lid). When the container 1001 is a closed-type container, the upper surface is also configured to be deformable so as not to hinder the deformation of the bottom 1010. Furthermore, when the container 1001 is a closed-type container, it is preferable to provide an opening in the upper surface or the second side wall 1022, etc., as a location for receiving a culture medium or the like from the outside. By making the container 1001 a closed-type container, it is possible to prevent the liquid such as the culture medium 1003 and / or CO 2 from being injected into the container 1001. ２ It is also possible to increase the internal pressure of the container 1001 by using a gas such as the above, thereby expanding and contracting the container 1001.

[0177] (First Modification of Second Embodiment) Next, a container 1001A according to a first modification of the second embodiment will be described with reference to Fig. 16. Fig. 16 is a diagram for explaining a container 1001A according to the first modification of the second embodiment.

[0178] As shown in FIG. 16, a container 1001A in this modification has a bottom 1010A and a sidewall 1020A, similar to the container 1001 in the second embodiment.

[0179] In this modified example, the bottom 1010A of the container 1001A is deformed according to the processing stage of the cell 1002, similar to the container 1001 in the second embodiment described above, but unlike the container 1001 in the second embodiment described above, the side wall 1020A is not deformed, and only the bottom 1010A is deformed.

[0180] In container 1001A according to this modification, bottom 1010A is made of an elastic material, similar to bottom 1010 in embodiment 2. Specifically, bottom 1010A is an elastic member having rubber elasticity, and is made of an elastomer such as silicone rubber. It is preferable that bottom 1010A has flexibility similar to that of a rubber balloon.

[0181] The side wall portion 1020A is made of a non-stretchable material. Specifically, the side wall portion 1020A is made of the same material as the second side wall portion 1022 in the second embodiment. For example, the side wall portion 1020A is made of a hard resin material or a metal material. The side wall portion 1020A may be in the shape of a rectangular tube or a cylindrical tube.

[0182] 16(a) and 16(b), when a liquid such as culture medium 1003 is added to container 1001A, bottom 1010A deforms so as to bulge downward. That is, bottom 1010A bulges downward due to the weight of the liquid such as culture medium 1003. For example, bottom 1010A deforms so as to curve into a dome shape.

[0183] Then, as the bottom 1010A of the container 1001A deforms, the area of ​​the bottom surface 1010a of the bottom 1010A changes. Specifically, as the bottom 1010A deforms so as to bulge, the area of ​​the bottom surface 1010a of the bottom 1010A increases. In other words, by adding culture medium 1003 to the container 1001A, the area of ​​the bottom surface 1010a of the bottom 1010A can be increased. This increases the volume of the container 1001A.

[0184] The container 1001A configured as described above is used when processing the cells 1002. Also in this modification, similarly to the second embodiment, the container 1001A is a culture container for culturing the cells 1002, and the container 1001A is used when culturing the cells 1002.

[0185] 16 , when cells 1002 are cultured, cells 1002 are supplied to container 1001A, and culture medium 1003 for culturing cells 1002 is also supplied. Similarly to container 1001 in the second embodiment, container 1001A has bottom 1010A that deforms depending on the stage of processing cells 1002. Specifically, in this modification, container 1001A also has a bottom surface 1010a that changes area depending on the culture stage.

[0186] As described above, in the container 1001A according to this modification, the bottom 1010A of the container 1001A is deformed depending on the processing stage, similar to the container 1001 according to the second embodiment.

[0187] This allows efficient processing of cells 1002 by container 1001A. For example, cells 1002 can be efficiently cultured and proliferated by deforming bottom 1010A of container 1001A according to the culture stage.

[0188] Moreover, in the container 1001A of this modification, only the bottom 1010A of the container 1001A is deformed by the weight of the culture medium 1003. This allows the bottom 1010A to be easily deformed without using a drive device, even in a case where the cell culture space is limited and it is not possible to deform the bottom 1010A using a drive device.

[0189] Also in this modification, the area of ​​bottom surface 1010a of bottom portion 1010A of container 1001A changes as bottom portion 1010A of container 1001A deforms. Specifically, the area of ​​bottom surface 1010a of bottom portion 1010A of container 1001A increases as bottom portion 1010A of container 1001A expands and deforms.

[0190] In this way, by increasing the area of ​​the bottom surface 1010a of the bottom portion 1010A, it is possible to efficiently carry out adhesion culture of the cells 1002. This allows the cells 1002 to be efficiently cultured and proliferated.

[0191] In this modification, the container 1001A is a cylindrical container with a bottom and an opening 1001a, but is not limited to this. For example, the container 1001A may be a closed-type container with an upper surface (top lid). In this case, an opening may be provided in the upper surface or the side wall 1020A as a place to receive a culture medium or the like from the outside. By making the container 1001A a closed-type container, liquid such as the culture medium 1003 and / or CO 2 to be injected into the container 1001A can be easily received. ２ It is also possible to increase the internal pressure of the container 1001A with a gas such as the above, thereby expanding and contracting the bottom 1010A of the container 1001A.

[0192] (Second Modification of Second Embodiment) Next, a container 1001B according to a second modification of the second embodiment will be described with reference to Fig. 17. Fig. 17 is a diagram for explaining a container 1001B according to the second modification of the second embodiment.

[0193] In this modified example, the bottom 1010B of the container 1001B is deformed according to the processing stage of the cell 1002, similar to the container 1001 in the second embodiment, but the overall shape of the container 1001B in this modified example is different from that of the container 1001 in the second embodiment.

[0194] 17, a container 1001B according to this modification has a bottom 1010B and a cover 1030 that covers the bottom 1010B. The bottom 1010B and the cover 1030 form the container 1001B in the shape of a long, narrow bag.

[0195] 17A, when no liquid such as a culture medium is contained in the container 1001B of this modified example, the bottom 1010B and the cover 1030 are closely joined together. In this modified example, the bottom 1010B and the cover 1030 are bonded together with an adhesive that is easily peeled off.

[0196] As shown in (b) and (c) of FIG. 17, the container 1001B contains a liquid such as a culture medium 1003 and / or CO ２ As a result of the injection of a gas such as liquid or gas into the container 1001B, the internal pressure of the container 1001B increases, and the bottom 1010B expands and deforms so that the area between the bottom 1010B and the cover 1030 increases. Specifically, as the liquid and gas are added to the container 1001B, the internal pressure of the container 1001B gradually increases, causing the adhesive between the bottom 1010B and the cover 1030 to peel off, and the bottom 1010B gradually expands and deforms so that the area between the bottom 1010B and the cover 1030 increases. Then, as shown in (c) of FIG. 17 , the container 1001B is finally deformed by the bottom 1010B and the cover 1030 to become an elongated bag-like housing.

[0197] 17(b) and 17(c), when a liquid is stored in the container 1001B, it is advisable to attach a cap 1040 to the opening 1001a of the container 1001B, thereby preventing the liquid stored in the container 1001B from leaking out of the container 1001B.

[0198] The bottom 1010B and the cover 1030 may be made of a stretchable material or a non-stretchable material. The bottom 1010B and the cover 1030 are made of, for example, a resin material, but are not limited to this.

[0199] The container 1001B configured as described above is used when processing the cells 1002. In this modification, the container 1001B is also a culture container for culturing the cells 1002, and the container 1001B is used when culturing the cells 1002.

[0200] 17 , when cells 1002 are cultured, cells 1002 are supplied to container 1001B, and culture medium 1003 for culturing cells 1002 is also supplied. Similarly to container 1001 in the second embodiment, container 1001B has bottom 1010B that deforms depending on the stage of processing cells 1002. Specifically, in this modification, container 1001B also has a bottom surface 1010a that changes area depending on the culture stage.

[0201] As described above, in the container 1001B according to this modification, the bottom 1010B of the container 1001B is deformed depending on the processing stage, similar to the container 1001 according to the second embodiment.

[0202] This allows efficient processing of cells 1002 by container 1001B. For example, by deforming bottom 1010B of container 1001B according to the culture stage, cells 1002 can be efficiently cultured and proliferated.

[0203] Also in this modification, the area of ​​the bottom surface 1010a of the bottom portion 1010B of the container 1001B changes as the bottom portion 1010B of the container 1001B deforms. Specifically, the area of ​​the bottom surface 1010a of the bottom portion 1010B of the container 1001B increases as the bottom portion 1010B of the container 1001B expands and deforms.

[0204] In this way, by increasing the area of ​​the bottom surface 1010a of the bottom portion 1010B, it is possible to efficiently carry out adhesion culture of the cells 1002. This allows the cells 1002 to be efficiently cultured and proliferated.

[0205] Furthermore, in the container 1001B of this modification, the bottom surface 1010a of the bottom 1010B of the container 1001B increases as the amount of culture medium 1003 contained in the container 1001B increases. This allows the cells 1002 to be efficiently cultured and proliferated by adding the culture medium 1003 to the container 1001B.

[0206] In the above-mentioned second embodiment and its variant example 1, the containers 1001 and 1001A are open-type containers, and when the cells 1002 are cultured, the openings 1001a of the containers 1001 and 1001A are open. However, in this variant example, the container 1001B is a closed-type container, and when the cells 1002 are cultured, the openings 1001a of the container 1001B are not open, and a cap 1040 is attached to the openings 1001a.

[0207] Furthermore, as shown in FIG. 18 , the container 1001B may have a scale 1050. The scale 1050 is provided on the bottom 1010B of the container 1001B. As shown in FIG. 19 , the scale 1050 has graduations drawn on it. As the container 1001B expands, the bottom 1010B expands into a planar shape, so that the graduations on the scale 1050 function as an original graduation. In other words, as the container 1001B expands and the cells 1002 proliferate, the scale 1050 functions as a graduation. This allows the density and the like of the proliferated cells 1002 to be measured by the scale 1050. In this way, by measuring the density and the like of the cells 1002 by the scale 1050, it becomes possible to appropriately determine the timing when expansion culture is necessary.

[0208] In this modification, bottom 1010B and cover 1030 are bonded together with an adhesive, but this is not a limitation. That is, bottom 1010B and cover 1030 do not have to be bonded together with an adhesive. In this case, container 1001B itself may memorize its shape in a curled state like a whistle, and container 1001B may expand and deform when liquid such as culture medium 1003 is poured into container 1001B, or container 1001B may contract and return to its curled state when liquid such as culture medium 1003 flows out of container 1001B.

[0209] (Third Modification of Second Embodiment) Next, a container 1001C according to a third modification of the second embodiment will be described with reference to Fig. 20. Fig. 20 is a diagram for explaining a container 1001C according to the third modification of the second embodiment.

[0210] As shown in FIG. 20, a container 1001C in this modification has a bottom 1010C and a sidewall 1020C, similar to the container 1001 in the second embodiment.

[0211] Like the container 1001 in the second embodiment, the container 1001C in this modification has a bottom 1010C that deforms depending on the stage of processing of the cells 1002, but the configuration of the bottom 1010C differs from that of the container 1001 in the second embodiment.

[0212] In the container 1001C according to this modification, the bottom 1010C has a bellows-shaped bellows portion 1011. In this modification, the bottom 1010C of the container 1001C is deformed as the bellows portion 1011 expands and contracts. The bellows portion 1011 can be made of any material that can deform into a bellows shape. For example, the bellows portion 1011 can be made of a resin material or a metal material. Furthermore, the material of the bellows portion 1011 itself may or may not be stretchable.

[0213] The bottom portion 1010C also has a side plate portion 1012. The side plate portion 1012 is connected to the side wall portion 1020C. The side plate portion 1012 moves in accordance with the deformation of the bellows portion 1011. Specifically, the side plate portion 1012 moves around the connection portion with the side wall portion 1020C of the container 1001C as a fulcrum. The side plate portion 1012 is made of an elastic material. Specifically, the side plate portion 1012 is an elastic member having rubber elasticity, and is made of an elastomer such as silicone rubber.

[0214] Unlike the second embodiment, the side wall portion 1020C does not deform. Therefore, the side wall portion 1020C is made of a material that does not expand or contract as a whole. Specifically, the side wall portion 1020C is made of the same material as the second side wall portion 1022 in the second embodiment. For example, the side wall portion 1020C is made of a hard resin material or a metal material. The side wall portion 1020C may be in the shape of a rectangular tube or a cylinder.

[0215] In container 1001C configured in this manner, as shown in (a) to (c) of Figure 20, when a liquid such as culture medium 1003 is added to container 1001C, bellows portion 1011 of bottom 1010C expands and bottom 1010C is deformed. Specifically, in (a) of Figure 20, bellows portion 1011 of bottom 1010C contracts and bottom 1010C is in a folded state, but when culture medium 1003 is added to container 1001C, bellows portion 1011 expands and folded bottom 1010C unfolds as shown in (b) of Figure 20, and when culture medium 1003 is further added to container 1001C, bellows portion 1011 unfolds into a flat shape and bottom 1010C further unfolds, so that the entire bottom 1010C becomes flat as shown in (c) of Figure 20.

[0216] Then, the bottom 1010C of the container 1001C is deformed as a result. Specifically, the bellows portion 1011 of the bottom 1010C expands, causing the bottom 1010C to expand and deform. In other words, as the culture medium 1003 is added to the container 1001C, the bottom surface 1010a of the bottom 1010C expands into a flat shape, and the volume of the container 1001C increases.

[0217] The container 1001C configured as above is used when processing cells. In this modification, the container 1001C is also a culture container for culturing cells, and the container 1001C is used when culturing cells.

[0218] 20 , when cells are cultured, cells (not shown) are supplied to container 1001C, along with culture medium 1003 for culturing the cells. Similarly to container 1001 in the second embodiment, container 1001C has bottom 1010C that deforms depending on the stage of cell processing.

[0219] As described above, in the container 1001C according to this modification, the bottom 1010C of the container 1001C is deformed depending on the processing stage, similar to the container 1001 according to the second embodiment.

[0220] This allows cells to be efficiently processed by container 1001 C. For example, by deforming bottom 1010 C of container 1001 C according to the culture stage, cells can be efficiently cultured and grown.

[0221] Moreover, in the container 1001C of this modified example, the bottom 1010C of the container 1001C is deformed by the weight of the culture medium 1003. This allows the bottom 1010C to be easily deformed without using a drive device, even in a case where the cell culture space is limited and the bottom 1010C cannot be deformed by a drive device.

[0222] In this modification, bottom 1010C of container 1001C has bellows-shaped bellows portion 1011, and as bellows portion 1011 expands, bottom 1010C of container 1001C expands and deforms.

[0223] This increases the area of ​​the flat portion of the bottom surface 1010a (the actual area available for adhesion culture), allowing efficient adhesion culture of cells, thereby enabling efficient cell culture and proliferation.

[0224] In this modification, bellows portion 1011 of bottom portion 1010C of container 1001C is folded multiple times when in the contracted state, but this is not limited to this. For example, as in container 1001D shown in Fig. 21 , bellows portion 1011D of bottom portion 1010D may be folded only once when in the contracted state. In this case, bellows portion 1011D may be made of an elastic material.

[0225] (Embodiment 3) Next, the configuration of a container 2001 according to embodiment 3 will be described with reference to Fig. 22 and Fig. 23. Fig. 22 is a perspective view of the container 2001 according to embodiment 3. Fig. 23 is a cross-sectional view of the container 2001 according to embodiment 3.

[0226] The container 2001 shown in Fig. 22 is a container for processing cells. In this embodiment, the container 2001 is a cell culture container for culturing cells. For example, the container 2001 processes human-derived cells such as iPS cells. As an example, the container 2001 cultures human stem cells, etc. Culturing the cells in the container 2001 causes the cells to proliferate.

[0227] The use of the container 2001 is not limited to cell culturing. For example, the container 2001 may be used to store cells or reagents, guide specific cells, or extract specific cells.

[0228] iPS cells are produced from hematopoietic stem cells contained in blood. Specifically, iPS cells can be obtained by infecting hematopoietic stem cells extracted from blood with a viral vector and then introducing an iPS cell transformation gene into the infected hematopoietic stem cells. The cells processed in container 2001 may be T cells obtained by further inducing differentiation of iPS cells, stem cells other than iPS cells such as ES cells, or cells other than stem cells.

[0229] The container 2001 in this embodiment is a closed container. That is, the container 2001 is a container that can be sealed, and cells can be treated in a sealed state of the container 2001. Therefore, by using the container 2001, cell culture can be performed in a closed system.

[0230] 23 , the container 2001 has a container body 2010 having an opening 2010a, and a protective layer 2020 provided on the inner surface of the container body 2010. The container 2001 further has a cap 2030 that closes the opening 2010a of the container body 2010. By closing the opening 2010a of the container body 2010 with the cap 2030, the container 2001 can be sealed, and the container 2001 can be made into a closed container. The cap 2030 is, for example, a resin cap made of resin or a metal cap made of metal.

[0231] The container body 2010 is an outer container that forms the outer shell of the container 2001. The container body 2010 is a light-transmitting case. In this embodiment, the container body 2010 is a transparent case so that the contents can be seen. The container body 2010 is made of a light-transmitting material such as a light-transmitting resin material or glass. The container body 2010 may be flexible or may be inflexible. In this embodiment, the container body 2010 is not flexible and has a fixed shape that does not change shape.

[0232] For example, the container body 2010 is made of a hard, transparent resin material. Specifically, the container body 2010 is made of a plant-derived resin (bio-resin) that is transparent and highly heat-resistant. More specifically, the container body 2010 is made of a biodegradable resin such as polylactic acid resin. Polylactic acid resin is a biodegradable resin made from polylactic acid obtained from starch such as corn or potato. In other words, the container body 2010 is made of renewable, plant-derived raw materials. In this way, by making the container body 2010 of a biodegradable resin such as polylactic acid resin, it is possible to impart biodegradability to the container body 2010 in addition to the performance of existing cell culture containers.

[0233] As shown in FIGS. 22 and 23, the container body 2010 has an upper wall portion 2011 , a bottom wall portion 2012 facing the upper wall portion 2011 , a pair of side wall portions 2013 and 2014 , a front wall portion 2015 , and a rear wall portion 2016 .

[0234] Each of the pair of side walls 2013 and 2014 has an inclined portion that slopes toward the front wall 2015. The front wall 2015 has a neck 2017 having an opening 2010a. The neck 2017 is a cylindrical portion. The inside of the container body 2010 can be accessed from the neck 2017. A threaded portion that threads onto the outer surface of the neck 2017 and the inner surface of the cap 2030 is formed, and the opening 2010a of the neck 2017 can be covered by screwing the cap 2030 onto the neck 2017.

[0235] In this embodiment, the cap 2030 is provided as a lid on the opening 2010a of the neck 2017. However, the present invention is not limited to this. For example, the neck 2017 may be configured so that a tube or the like leading to another closed container can be connected thereto.

[0236] A protective layer 2020 is provided on the inner surface of the container body 2010. In this embodiment, the protective layer 2020 is provided on the entire inner surface of the container body 2010. Specifically, the protective layer 2020 is provided continuously over the top wall portion 2011, the bottom wall portion 2012, the side wall portion 2013, the side wall portion 2014, the front wall portion 2015, the rear wall portion 2016, and the neck portion 2017. The inner surface of the protective layer 2020 is the inner surface of the container 2001. Therefore, cells, fluid, etc. supplied to the container 2001 come into contact with the protective layer 2020.

[0237] The protective layer 2020 is fixed to the inner surface of the container body 2010. Specifically, the protective layer 2020 is adhered and fixed to the inner surface of the container body 2010. The protective layer 2020 is made of, for example, a resin material. By adhering and fixing the protective layer 2020 to the inner surface of the container body 2010, it is possible to provide a stable environment when processing cells, particularly when culturing cells.

[0238] The protective layer 2020 may be a film adhered to the inner surface of the container body 2010, or may be a coating film adhered by coating the inner surface of the container body 2010. When the protective layer 2020 is a film, an adhesive layer made of an adhesive or the like is present at the interface between the protective layer 2020 and the container body 2010. When the protective layer 2020 is a coating film, the protective layer 2020 may be directly bonded to the container body 2010, or the protective layer 2020 and the container body 2010 may be bonded via an adhesive layer.

[0239] In this embodiment, the protective layer 2020 is a film having an adhesive layer. In this case, for example, as shown in FIG. 24( a), the protective layer 2020, which is a bag-shaped film with an adhesive applied to its outer surface, is inserted into the container body 2010 through the opening 2010a. Next, as shown in FIG. 24( b), a fluid supply nozzle 2003 is attached to the neck portion 2017 of the container body 2010, and a fluid 2002, such as a liquid or gas, is supplied from the fluid supply nozzle 2003 into the container body 2010. Specifically, the fluid 2002 is supplied from the fluid supply nozzle 2003 into the bag-shaped protective layer 2020. As a result, the bag-shaped protective layer 2020 expands and becomes larger, and the protective layer 2020 adheres to the container body 2010. In other words, the adhesive applied to the outer surface of the protective layer 2020 adheres to the inner surface of the container body 2010, thereby adhering the protective layer 2020 to the container body 2010. Thereafter, although not shown, the fluid supply nozzle 2003 is removed from the container body 2010 and the fluid 2002 inside the container body 2010 is discharged, thereby obtaining the container 2001 shown in FIG.

[0240] In the present embodiment, a film coated with adhesive is used as the protective layer 2020, and the protective layer 2020 is inserted into the container body 2010 to adhere to the inner surface of the container body 2010. However, this is not limiting. For example, an adhesive may be applied to the inner surface of the container body 2010 in advance, and a film not coated with adhesive may be used as the protective layer 2020. The protective layer 2020 may be inserted into the container body 2010, and the fluid 2002 may be supplied into the container body 2010 to expand the protective layer 2020, thereby adhering the protective layer 2020 to the inner surface of the container body 2010. Alternatively, the protective layer 2020 not coated with adhesive may be inserted into the container body 2010 to bring the protective layer 2020 into close contact with the inner surface of the container body 2010, and then the protective layer 2020 may be adhered to the container body 2010 by pouring an adhesive therein or irradiating it with ultraviolet (UV) light.

[0241] The container 2001 thus produced is used, for example, to process cells. In this case, cells are placed inside the container 2001 and a predetermined process is performed. After the cells have been processed, the used container 2001 has contaminated inner surfaces. Specifically, as shown in FIG. 25( a), the used container 2001 has contaminants 2004 adhering to the inner surface of the protective layer 2020.

[0242] When disposing of used container 2001 having contaminants 2004 attached thereto, it is necessary for the user to avoid contact with contaminants 2004. For example, when blood is poured into container 2001, blood adheres to the inner surface of protective layer 2020 as contaminants 2004. If the user comes into contact with the blood, there is a risk of developing an infectious disease, so when disposing of used container 2001, it is necessary for the user to avoid contact with contaminants 2004.

[0243] Therefore, in this embodiment, when disposing of a used container 2001, as shown in (b) of FIG. 25 , the contaminants 2004 inside the container 2001 are removed by peeling the protective layer 2020 to which the contaminants 2004 are attached from the container body 2010. Specifically, the cap 2030 is held and the base of the neck 2017 is snapped off to separate the cap 2030 from the container body 2010, and the protective layer 2020 connected to the cap 2030 is physically peeled off from the container body 2010 by pulling the cap 2030, thereby pulling the protective layer 2020 from the container body 2010. The protective layer 2020 pulled out from the container body 2010 can be rolled up and discarded. In other words, of the container body 2010 and the protective layer 2020, only the protective layer 2020 to which the contaminants 2004 are attached can be discarded. In this case, it is advisable to design neck portion 2017 to be two to three times longer than cap 2030. This allows container body 2010 to be recovered by breaking neck portion 2017 every time container 2001 is used, so that container body 2010 can be used repeatedly.

[0244] Instead of separating the cap 2030 from the container body 2010 by cutting the neck portion 2017, the cap 2030 may be turned to remove it from the neck portion 2017 and then separated from the container body 2010. In this case, the protective layer 2020 may be formed by folding it back at the opening 2010a, so that the protective layer 2020 extends to the outer surface of the neck portion 2017. In other words, the protective layer 2020 may also be adhered to the inner surface of the cap 2030. This allows the cap 2030 to be easily peeled off from the container body 2010 together with the protective layer 2020 adhered to the cap 2030 by pulling the cap 2030 after turning the cap 2030 to remove it from the neck portion 2017. Furthermore, by turning the cap 2030, the protective layer 2020 adhered to the cap 2030 can also be easily peeled off from the container body 2010 by the rotational force.

[0245] Furthermore, in this embodiment, instead of discarding only the protective layer 2020, the cap 2030 is pulled to separate the protective layer 2020 and the cap 2030 from the container body 2010. As a result, even if contaminants 2004 are attached not only to the protective layer 2020 but also to the cap 2030, the cap 2030 with the contaminants 2004 attached can be discarded together with the protective layer 2020 with the contaminants 2004 attached. In other words, the cap 2030 can be discarded together with the protective layer 2020.

[0246] As described above, the container 2001 of this embodiment is a closed container for processing cells, and has a container body 2010 and a protective layer 2020 provided on the inner surface of the container body 2010, and the protective layer 2020 is configured so that it can be peeled off from the container body 2010 by physical means.

[0247] With this configuration, by peeling the protective layer 2020 from the container body 2010, it is possible to remove the areas in the container 2001 that have come into contact with the contaminants 2004. This allows the used container 2001 to be disposed of without touching the areas in the container 2001 that have come into contact with the contaminants 2004. Furthermore, by peeling the protective layer 2020, the contaminants 2004 are no longer present in the container body 2010, so that the container body 2010 from which the protective layer 2020 has been peeled can be re-formed with the protective layer 2020 and used as a new container 2001. This allows the container body 2010 to be recycled while preventing contamination, thereby suppressing adverse effects on the environment.

[0248] In particular, in this embodiment, the container body 2010 is biodegradable. This allows the container body 2010 to be decomposed into carbon dioxide and water by microorganisms, and therefore does not have a negative impact on the environment. In other words, an environmentally friendly container 2001 can be realized.

[0249] In this embodiment, when peeling the protective layer 2020 from the container body 2010, the protective layer 2020 is peeled off from the container body 2010 by pulling out the protective layer 2020, but the present invention is not limited to this.

[0250] For example, as shown in (a) of FIG. 26 , a used container 2001 having contaminants 2004 adhering to the inner surface of the protective layer 2020 may be subjected to heat treatment to peel the protective layer 2020 from the container body 2010. In this case, when the used container 2001 is subjected to heat treatment, the protective layer 2020 shrinks due to the heat and peels off from the container body 2010, as shown in (b) of FIG. 26 . Then, by removing the cap 2030 and removing the shrunken protective layer 2020 from the opening 2010a of the container body 2010, only the protective layer 2020 with the contaminants 2004 adhering thereto can be discarded. In this modification, the protective layer 2020 is heated and hardened, making it difficult for a user to come into contact with the contaminants 2004 when discarding the protective layer 2020. In this modification, the protective layer 2020 is made of a material that hardens when heated. In this modification, the cap 2030 is removed after the heat treatment, but this is not limited to this. Specifically, the cap 2030 may be removed and then heat treatment may be performed to peel off the protective layer 2020 .

[0251] In the above embodiment, the protective layer 2020 is peeled off from the container body 2010 by a physical method, but the present invention is not limited to this. Specifically, the protective layer 2020 may be peeled off from the container body 2010 by a chemical method.

[0252] For example, as shown in (a) of FIG. 27 , ultraviolet (UV) light can be applied to a used container 2001 having contaminants 2004 attached to the inner surface of the protective layer 2020, thereby peeling the protective layer 2020 from the container body 2010. In this case, when the used container 2001 is irradiated with ultraviolet light, the adhesive layer at the interface between the protective layer 2020 and the container body 2010 loses its adhesiveness, causing the protective layer 2020 to peel off (for example, the adhesive dries and peels off, or the materials of the protective layer 2020 and the adhesive layer deteriorate, resulting in a difference in elasticity between them and causing peeling). As shown in (b) of FIG. 27 , the protective layer 2020 floats away from the container body 2010. As a result, the protective layer 2020 peels off from the container body 2010. Therefore, in this modification, it is preferable that the adhesive layer at the interface between the protective layer 2020 and the container body 2010 be made of a material that peels off when exposed to ultraviolet light.

[0253] By irradiating the container 2001 with ultraviolet light in this manner, not only can the protective layer 2020 be peeled off from the container body 2010, but also the ultraviolet light can sterilize the contaminants 2004 inside the container 2001. Therefore, the peeled protective layer 2020 can be safely disposed of.

[0254] Although not shown, the peeled protective layer 2020 may be discarded by removing it from the opening 2010a of the container body 2010, or as shown in Figure 28, the protective layer 2020 may be discarded by dissolving it with a liquid chemical 2005 such as a solvent.

[0255] (Modification of Third Embodiment) Next, a container 2001A according to a modification of the third embodiment will be described with reference to Fig. 29. Fig. 29 is a diagram showing the configuration of a container 2001A according to a modification of the third embodiment.

[0256] 29, a container 2001A according to this modification has a structure in which a pipe 2018 is provided in a part of the container 2001A, as compared to the container 2001 according to the third embodiment. The other configurations are the same as those of the third embodiment.

[0257] The piping 2018 is provided in the container body 2010A. In this modification, the piping 2018 is provided in the side wall portion 2013 of the container body 2010A. Specifically, the piping 2018 is provided in the inclined portion of the side wall portion 2013.

[0258] The piping 2018 has an opening 2019 that leads between the container body 2010A and the protective layer 2020. That is, when the protective layer 2020 is formed on the container body 2010A, the opening 2019 is blocked by the protective layer 2020, and the protective layer 2020 can be accessed via the piping 2018. Note that when the protective layer 2020 is not formed on the container body 2010A, the internal space of the container body 2010A communicates with the outside air via the piping 2018, but when the protective layer 2020 is formed on the container body 2010A, the internal space of the container body 2010A is no longer in communication with the outside air.

[0259] The protective layer 2020 is, for example, a film having an adhesive layer on its outer surface. In this case, for example, as shown in FIG. 30(a), the protective layer 2020, which is a bag-shaped film with an adhesive applied to its outer surface, is inserted into the container body 2010A through the opening 2010a, and the opening 2010a is closed with the cap 2030. Next, as shown in FIG. 30(b), the protective layer 2020 is inflated by suctioning air from the container body 2010A through the piping 2018. This causes the inflated protective layer 2020 to adhere to the container body 2010A. In other words, the adhesive applied to the outer surface of the protective layer 2020 adheres to the inner surface of the container body 2010A, thereby adhering the protective layer 2020 to the container body 2010A. In this manner, the container 2001A can be obtained.

[0260] In this modified example, instead of inserting the protective layer 2020 coated with adhesive into the container body 2010A, adhesive may be applied to the inner surface of the container body 2010A in advance, and the protective layer 2020 without adhesive may be inserted into the container body 2010A to expand the protective layer 2020 and adhere to the inner surface of the container body 2010A. Alternatively, the protective layer 2020 without adhesive may be inserted into the container body 2010A and the protective layer 2020 may be closely attached to the inner surface of the container body 2010A, and then adhesive may be poured in or ultraviolet (UV) light may be irradiated to adhere the protective layer 2020 to the container body 2010A.

[0261] The container 2001A thus obtained is then used to process cells in the same manner as in the above-described embodiment 3. In the used container 2001A after cell processing, contaminants 2004 are attached to the inner surface of the protective layer 2020, as shown in Figure 31(a).

[0262] In this modification, when disposing of a used container 2001A having contaminants 2004 attached thereto, the contaminants 2004 are removed from the container body 2010A by peeling the protective layer 2020 to which the contaminants 2004 are attached from the container body 2010A. Specifically, as shown in FIG. 31 (b), a liquid 2006 is supplied from a pipe 2018 to be introduced between the container body 2010A and the protective layer 2020. As the liquid 2006, water can be used if the adhesive is water-soluble, or a specific dissolving liquid (e.g., a liquid that dissolves the adhesive by chemical reaction) can be used if the adhesive is to be dissolved. In this manner, by supplying the liquid 2006, such as water, between the container body 2010A and the protective layer 2020, the adhesive layer at the interface between the container body 2010A and the protective layer 2020 peels off, allowing the protective layer 2020 to float from the container body 2010A. That is, the protective layer 2020 is physically peeled off from the container body 2010A. Thereafter, the protective layer 2020 peeled off from the container body 2010A can be pulled out from the container body 2010A and discarded. That is, in this modification as well, only the protective layer 2020 to which the contaminants 2004 are attached can be discarded.

[0263] As described above, the container 2001A of this modified example is a closed container for processing cells, similar to the third embodiment above, and has a container body 2010A and a protective layer 2020 provided on the inner surface of the container body 2010A, and the protective layer 2020 is configured so that it can be peeled off from the container body 2010A by physical means.

[0264] With this configuration, by peeling the protective layer 2020 from the container body 2010A, it is possible to remove the areas in the container 2001A that have come into contact with the contaminant 2004. This allows the used container 2001A to be disposed of without touching the areas in the container 2001A that have come into contact with the contaminant 2004. Furthermore, because the contaminant 2004 is no longer present in the container body 2010A by peeling the protective layer 2020, the container body 2010A from which the protective layer 2020 has been peeled can be re-formed with the protective layer 2020 and used as a new container 2001A. This allows the container body 2010 to be recycled while preventing contamination, thereby suppressing adverse effects on the environment.

[0265] In this modification, the protective layer 2020 may also be peeled off from the container body 2010A by a chemical method, rather than by a physical method.

[0266] In addition, in this modification, the piping 2018 is provided on the side wall portion 2013 of the container body 2010A, but this is not limiting. For example, the piping 2018 may be provided at a location other than the side wall portion 2013 of the container body 2010A.

[0267] Furthermore, in this modification, the opening 2019 is provided in the container body 2010A by providing the piping 2018, but this is not limiting. For example, as in the container 2001B shown in FIG. 32 , the opening 2019 may be provided in the container body 2010B without providing the piping. Note that in FIG. 32 , the opening 2019 is provided in the side wall portion 2013 of the container body 2010B, but this is not limiting. For example, the opening 2019 may be provided in a location other than the side wall portion 2013 of the container body 2010B.

[0268] After cells have been treated using container 2001B configured in this manner, the used container 2001B to which contaminants 2004 have adhered can be disposed of as shown in Fig. 33. Specifically, as shown in Fig. 33, by immersing container 2001B in a container 2100 such as a tub containing liquid 2007 such as water, liquid 2007 penetrates and penetrates between container body 2010B and protective layer 2020, causing protective layer 2020 to float up from container body 2010B. This allows protective layer 2020 to be physically peeled off from container body 2010B.

[0269] (Other Modifications) The technology of the present disclosure has been described above based on the first to third embodiments and their modifications, but the present disclosure is not limited to the above-described first to third embodiments and their modifications.

[0270] For example, in the first embodiment, iPS cells are produced from hematopoietic stem cells using a viral vector, but the present invention is not limited to this. In other words, iPS cells may be produced from hematopoietic stem cells without using a viral vector.

[0271] Furthermore, in the first embodiment, the cell culture device 100 and the cell generation device 100A are closed types, and the cell culture method and the cell generation method are closed systems, but this is not limited to this. The technology of the present disclosure may be applied to open-type cell culture devices and cell generation devices, or to open-system cell culture methods and cell generation methods. However, the technology of the present disclosure is suitable for closed-type cell culture devices and cell generation devices, and closed-system cell culture methods and cell generation methods.

[0272] In addition, in the second embodiment, the bottom of the container is deformed at the culture stage when cells are transitioned from reprogramming culture to expansion culture, but this is not limiting. For example, the bottom of the container may be deformed at the culture stage when cells are transitioned from infection to reprogramming culture, or at a culture stage at any other timing.

[0273] Furthermore, in the second embodiment, the cells 1002 to be treated in the container are iPS cells, but this is not limiting. For example, the cells 1002 to be treated in the container may be T cells obtained by further inducing differentiation of iPS cells, stem cells other than iPS cells such as ES cells, or cells other than stem cells.

[0274] In the third embodiment, the contaminants 2004 are attached to the entire inner surface of the protective layer 2020 in the used containers 2001 to 2001B, but this is not limiting. Specifically, the contaminants 2004 may be attached to only a portion of the inner surface of the protective layer 2020.

[0275] In addition, the present disclosure also includes forms obtained by applying various modifications that a person skilled in the art would conceive of to the above-mentioned first to third embodiments and their variations, and forms realized by arbitrarily combining the components and functions of each embodiment, etc., within the scope of the present disclosure. Furthermore, the present disclosure also includes any combination of two or more claims from among the multiple claims set forth in the claims at the time of filing, within the scope of technical compatibility. For example, when a dependent claim set forth in the claims at the time of filing is made into a multiple claim or multiple multiple claims that cite all of the superordinate claims within the scope of technical compatibility, the present disclosure also includes all combinations of claims included in that multiple claim or multiple multiple multiple claims.

[0276] The technology disclosed herein is useful as a cell culture method for culturing or generating cells such as iPS cells, a cell culture device, and a piping sheet used in the cell culture device.

[0277] Furthermore, the technology of the present disclosure is useful as a container for processing cells such as iPS cells.

[0278] 1, 1A Piping sheet 2 Centrifuge 3 Propeller 4 Mixing channel 10 Sheet body 10a First end 10b Second end 20 Piping 21 First pipe 21a First liquid pipe 21b First gas pipe 22 Second pipe 22a Second liquid pipe 22b Second gas pipe 23 Third pipe 23a Third liquid pipe 23b Third gas pipe 24 Fourth pipe 24a Fourth liquid pipe 24b Fourth gas pipe 25 Fifth pipe 25a Fifth liquid pipe 25b Fifth gas pipe 26, 27, 28, 29 Connecting pipe 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 30a, 30b, 30c, 30d, 30e First joint 40, 41, 42, 43, 44, 45, 40a, 40b, 40c, 40d Second joint 100 Cell culture device 100A Cell generation device 111 Cell container 112 Bead container 113 First culture medium container 114 Viral vector container 115 Second culture medium container 116 Third culture medium container 117 Fourth culture medium container 118 Differentiation induction factor container 119 Fifth culture medium container 121 Extraction container 122 Infection container 123 Culture container 124 Storage container 125 Disease mutation container 125a Container body 125b Cell compartment 125c, 125d Through-hole 126 Differentiation induction container 127 Storage container 200, 200A, 200B, 200C Storage container 210, 210B, 210C First space 220, 220B, 220C Second space 230 Communication path 310 Inducer introducer 320 Cell extractor 400 Observation device 1001, 1001A, 1001B, 1001C, 1001D Container 1001a Opening 1002 Cell 1003 Culture medium 1010, 1010A, 1010B, 1010C, 1010D Bottom 1010a Bottom surface 1011, 1011D Bellows portion 1012 Side plate portion 1020, 1020A, 1020C Side wall portion 1021 First side wall portion 1022 Second side wall portion 1030 Cover portion 1040 Cap 1050 Scale 2001, 2001A, 2001B Container 2002 Fluid 2003 Fluid supply nozzle 2004 Contaminant 2005 Chemical2006, 2007 Liquid 2010, 2010A, 2010B Container Body 2010a Opening 2011 Upper Wall 2012 Bottom Wall 2013, 2014 Side Wall 2015 Front Wall 2016 Rear Wall 2017 Head 2018 Piping 2019 Opening 2020 Protective Layer 2030 Cap 2100 Containers V1, V2, V3, V4, V5, V6, V7, V8, V9, V10, V11, V12, V13 Valve P, P1, P2, P3, P4, P5 Pump

Claims

1. A piping sheet used in carrying out a cell culture method including a first step and a second step, comprising: a sheet-shaped sheet body; a first pipe provided inside the sheet body for carrying out the first step; a second pipe provided inside the sheet body for carrying out the second step; a first joint provided at one open end of the first pipe and one open end of the second pipe, each of which is detachably connected to another member; and a second joint provided at the other open end of the first pipe and the other open end of the second pipe, each of which is detachably connected to another member, wherein the first joint of the first pipe and the first joint of the second pipe are each located at a first end of the sheet body, and the second joint of the first pipe and the second pipe are each located at a second end of the sheet body.

2. The piping sheet according to claim 1, wherein the second end portion is located opposite the first end portion.

3. The piping sheet according to claim 1, wherein the piping sheet is formed by bonding two sheets of sheet material together.

4. The piping sheet according to claim 1, wherein the first piping and the second piping are independent and do not merge.

5. A piping sheet as described in claim 1, wherein at least one of the first piping and the second piping includes a piping composed of a plurality of tributary pipings and a junction piping where the plurality of tributary pipings join together, the first joint is provided on each of the plurality of tributary pipings, and the second joint is provided on the junction piping.

6. The piping sheet according to claim 1, further comprising a connecting pipe provided inside the sheet body and connected to the second joint provided on the first pipe and the second joint provided on the second pipe.

7. A piping sheet according to any one of claims 1 to 6, wherein the first joint and the second joint are one-touch joints.

8. A piping sheet according to any one of claims 1 to 6, wherein the other component is any one of a liquid storage container that stores a predetermined liquid to be supplied to the piping sheet, a liquid recovery container that recovers liquid discharged from the piping sheet, a pump, and a valve.

9. A closed storage container for storing cells cultured using the piping sheet according to any one of claims 1 to 6, the storage container comprising a first space into which a liquid containing the cells flows, and a plurality of second spaces branching off from the first space.

10. The storage container according to claim 9, wherein two adjacent second spaces among the plurality of second spaces are spatially connected by a communication passage.

11. A cell generation system for carrying out a cell generation method including a mutation step of generating mutated iPS cells by introducing an induction factor into a portion of cultured iPS cells to cause mutation, and a differentiation induction step of mixing another portion of the cultured iPS cells with the mutated iPS cells and inducing differentiation, the cell generation system comprising a single closed processing device for carrying out the mutation step and the differentiation induction step.

12. The cell production system according to claim 11, wherein the processing device is the piping sheet according to claim 1.

13. The cell production system according to claim 11 or 12, comprising a closed processing vessel connected to the processing device for carrying out the mutation step.

14. A processing vessel used in generating mutant iPS cells by introducing an induction factor into cultured iPS cells, comprising: a sealed vessel body; and a plurality of cell compartments installed inside the vessel body, wherein the vessel body is configured so that the iPS cells set in the plurality of cell compartments can be observed from the outside, the vessel body is configured so that an induction factor introducer that introduces induction factors into the plurality of cell compartments can access the inside of the vessel body, and the vessel body is configured so that a cell extractor that extracts iPS cells from the plurality of cell compartments can access the inside of the vessel body.

15. The processing vessel according to claim 14, wherein the iPS cells set in the plurality of cell compartments are cultured using the piping sheet according to claim 1.

16. A container for processing cells, wherein the bottom of the container deforms according to the processing step.

17. The container according to claim 16, wherein the bottom of the container is deformed to change the area of ​​the bottom surface of the bottom of the container.

18. The container according to claim 16, wherein the bottom of the container has a bellows-shaped bellows portion, and the bottom of the container expands and deforms as the bellows portion expands.

19. The container according to any one of claims 16 to 18, wherein the treatment step is a culture step of culturing the cells.

20. The container according to claim 19, wherein the culture stage is a transition from an initiation culture to an expansion culture, or a transition from infection of cells to an initiation culture.

21. The container of claim 20, wherein the cells are human stem cells.

22. The container of claim 21, wherein the container is a closed container.

23. A closed container for processing cells, comprising: a container body; and a protective layer adhered to the inner surface of the container body, wherein the protective layer can be peeled off from the container body by a physical or chemical method.

24. The container according to claim 23, wherein the protective layer is a coating or film.

25. The container of claim 24, wherein the cells are of human origin.

26. The container according to claim 25, wherein the container body is biodegradable.

27. The container according to any one of claims 23 to 26, wherein the protective layer is peeled off from the container body by irradiating the container with ultraviolet light.

28. The container according to any one of claims 23 to 26, wherein the protective layer is peeled off from the container body by heating the container.

29. A container according to any one of claims 23 to 26, wherein an opening is provided in a part of the container that communicates between the container body and the protective layer.

30. The container according to claim 29, wherein the protective layer is peeled off from the container body by introducing a liquid between the container body and the protective layer through the opening.

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