Cell organization reactor

Abstract

Provided is a cell organization reactor with which it becomes possible to produce a cell population having an organized structure such as a biological tissue. A cell organization reactor 10 comprises a culture vessel 20, a culture medium container 30, and a control unit 40. The culture vessel 20 accommodates a cell sheet laminate 50 in which a first cell sheet 52 and a second cell sheet 54 are laminated on each other. A culture medium stored in the culture medium container 30 is supplied to the cell sheet laminate 50 in the culture vessel 20 via a culture medium intake pipe 60.

Description

Cell organization reactor

[0001] The present invention relates to a cell organization reactor.

[0002] A cell culture system for supplying a medium to cells and culturing the cells is known (see Patent Document 1).

[0003] Japanese Patent Application Laid-Open No. 2020-182392

[0004] In a conventional cell culture system, it has been difficult to culture a cell population having an organized structure such as a living tissue. The present invention has been made in view of the above problems, and an object thereof is to provide a cell organization reactor capable of producing a cell population having an organized structure such as a living tissue.

[0005] One aspect of the present invention is a cell organization reactor. The cell organization reactor includes a medium supply unit that supplies a medium to a cell sheet laminate in which a first cell sheet containing a specific type of cell and a second cell sheet containing a specific type of cell different from the specific type are laminated on at least one surface of the first cell sheet. In the cell organization reactor of the above aspect, the first cell sheet includes one or more cells selected from the group consisting of muscle-derived cells, skin-derived cells, intestine-derived cells, heart-derived cells, brain-derived cells, stomach-derived cells, embryonic membrane-derived cells, liver-derived cells, kidney-derived cells, and lung-derived cells, and the second cell sheet may include cells other than the cells used for the first cell sheet.

[0006] The cell organization reactor further includes a holding mechanism including a pair of holding parts that hold the cell sheet laminate in a sandwiched state, and the holding mechanism may gradually increase the distance between the pair of holding parts during the cell culture process by supplying the medium by the medium supply unit.

[0007] The cell sheet may be in a state where cells are seeded on a porous substrate.

[0008] The cell organization reactor may include a housing member that houses the cell sheet laminate and the pair of holding parts. The housing member may be stretchable according to the distance between the pair of holding parts.

[0009] According to the present invention, we can provide a technology relating to a cell organizing reactor that can produce a cell population having an organized structure similar to that of biological tissue.

[0010] Figure 1 shows an overview of the cell-organized reactor according to Embodiment 1. Figure 2 shows an overview of the cell-organized reactor according to Embodiment 2. Figure 3 shows a modified example of the holding structure of the cell sheet laminate.

[0011] Embodiments of the present invention will be described in detail below. In this specification, unless otherwise specified, the notation "a to b" in the description of numerical ranges means a or greater and b or less.

[0012] The cell assembly reactor according to the embodiment includes a culture medium supply unit that supplies culture medium to a cell sheet laminate, which is formed by laminating a first cell sheet containing a specific type of cell and a second cell sheet containing a different specific type of cell, which is arranged on at least one side of the first cell sheet. (Embodiment 1) Figure 1 is a diagram showing an overview of the cell assembly reactor 10 according to Embodiment 1. As shown in Figure 1, the cell assembly reactor 10 includes a culture vessel 20, a culture medium vessel 30, and a control unit 40.

[0013] The culture vessel 20 contains a cell sheet laminate 50. The cell sheet laminate 50 is a laminate formed by stacking a first cell sheet 52 and a second cell sheet 54.

[0014] The first cell sheet 52 contains a specific type of cell. The specific type of cell A includes one or more cells selected from the group consisting of muscle-derived cells, skin-derived cells, intestinal-derived cells, heart-derived cells, brain-derived cells, stomach-derived cells, germinal membrane-derived cells, liver-derived cells, kidney-derived cells, and lung-derived cells. The first cell sheet 52 is formed into a flat plate shape. The shape of the first cell sheet 52 when viewed from above is not particularly limited, but for example, it can be circular. The thickness of the first cell sheet 52 is not particularly limited, but from the viewpoint of improving the flowability of the culture medium, it is preferably 20 to 2000 μm, and more preferably 125 to 750 μm.

[0015] The second cell sheet 54 includes a specific type of cell B that is different from the specific type of cell described above. The specific type of cell B includes cells of the specific type of cell A (in other words, cells other than those used in the first cell sheet 52). For example, if the first cell sheet 52 is made of muscle-derived cells, the second cell sheet 54 includes vascular endothelium. Also, if the first cell sheet 52 is made of skin-derived cells, the second cell sheet 54 includes vascular endothelium. The second cell sheet 54 is laminated on one side of the first cell sheet. The second cell sheet 54 is formed into a flat plate shape. When viewed from above, the shape of the second cell sheet 54 is the same as that of the first cell sheet 52, and it is preferable that the first cell sheet 52 and the second cell sheet 54 overlap each other without overhanging. The thickness of the second cell sheet 54 is not particularly limited, but from the viewpoint of improving the flowability of the culture medium, it is preferably 20 to 2000 μm, and more preferably 125 to 750 μm. In this embodiment, the cell sheet laminate 50 has a two-layer structure in which the second cell sheet 54 is laminated on one side of the first cell sheet 52, but it may also have a three-layer structure in which the second cell sheet 54 is laminated on both sides of the first cell sheet 52.

[0016] The first cell sheet 52 and the second cell sheet 54 can be made from a porous substrate in which cells have been seeded. Examples of materials for the porous substrate include synthetic polymer materials and natural polymer materials. Examples of synthetic polymer materials include polyglycolic acid (PGA), polylactic acid (PLA), copolymer of polyglycolic acid and polylactic acid (PLGA), polyethylene glycol (PEG), and polycaprolactone. Examples of natural polymer materials include components of the extracellular matrix, specifically collagen, gelatin, alginic acid, hyaluronic acid, agarose, chitosan, fibrin, and fibroin. Polyethylene terephthalate and collagen are examples, and these can be formed into fibers or sheets and then molded into nonwoven fabrics, porous films, or meshes.

[0017] The cell sheet laminate 50 is held between a pair of holding members 56 and 58. By holding the cell sheet laminate 50 with the holding members 56 and 58, the cell sheet laminate 50 can be maintained so as not to collapse in shape during the initial stages of culture and throughout the culture process.

[0018] A predetermined culture medium is stored in the culture medium container 30. A well-known culture medium can be used depending on the cells to be cultured.

[0019] The culture medium container 30 and the culture vessel 20 are connected by a culture medium introduction tube 60 to allow liquid flow. The culture medium stored in the culture medium container 30 is introduced into the culture vessel 20 via the culture medium introduction tube 60. The culture medium introduced into the culture vessel 20 is supplied sequentially to the first cell sheet 52 and then to the second cell sheet 54. In other words, the culture medium supplied to the first cell sheet 52 passes through the first cell sheet 52 and is used to culture the cells contained in the first cell sheet 52, and then passes through the second cell sheet 54 and is used to culture the cells contained in the second cell sheet 54.

[0020] A pump 62 is installed in the culture medium introduction tube 60 for supplying a predetermined flow rate of culture medium to the culture vessel 20. The operation of the pump 62 is controlled by the control unit 40. The control unit 40 is not particularly limited as long as it can control the operation of the pump 62, but can be implemented, for example, by a computer equipped with an operation unit (not shown) and an input / output unit (not shown).

[0021] Furthermore, the culture medium container 30 and the culture vessel 20 are connected by a culture medium discharge pipe 64 to allow liquid flow. The culture medium that has passed through the second cell sheet 54 is returned to the culture medium container 30 via the culture medium discharge pipe 64.

[0022] According to the cell organizing reactor 10, by supplying culture medium to a cell sheet laminate 50 in which a first cell sheet 52 and a second cell sheet 54 are stacked, and performing cell culture, the cells contained in the first cell sheet 52 and the cells contained in the second cell sheet 54 become organized, thereby creating a cell population with an organized structure similar to biological tissue. Specifically, when the first cell sheet 52 and the second cell sheet 54 are formed from muscle-derived cells and vascular endothelial cells, respectively, a cell population organized like muscle can be obtained by culturing with the cell organizing reactor 10.

[0023] (Embodiment 2) Figure 2 is a diagram showing an overview of the cell organizing reactor 10 according to Embodiment 2. The basic configuration of the cell organizing reactor 10 according to Embodiment 2 is the same as that of Embodiment 1. Hereinafter, regarding the cell organizing reactor 10 according to Embodiment 2, explanations of the same configuration as Embodiment 1 will be omitted as appropriate, and configurations that differ from Embodiment 1 will be described.

[0024] In this embodiment, the distance between the holding member 56 and the holding member 58 is adjusted by a holding mechanism (not shown), such as an actuator. That is, the holding mechanism includes a pair of holding parts that hold the cell sheets stacked and sandwiched together. The operation of the holding mechanism is controlled by the control unit 40. Specifically, during the cell culture process, the distance between the holding member 56 and the holding member 58 is gradually increased. The control of the distance between the holding member 56 and the holding member 58 may be performed at a predetermined separation speed. In this case, the predetermined separation speed may be constant throughout the culture process, but the separation speed may be changed stepwise or continuously between the initial stage of culture and the stage after the initial stage of culture. Furthermore, the control of the distance between the holding member 56 and the holding member 58 may be feedback controlled while monitoring the cell culture conditions. For example, the pH of the culture medium may be monitored, and the separation speed may be changed according to the measured pH.

[0025] According to this embodiment, a group of cells having an organized structure like biological tissue can be grown according to the distance between the holding member 56 and the holding member 58.

[0026] In this embodiment, it is preferable that the culture container 20 (container) has a structure that can expand and contract according to the distance between the retaining member 56 and the retaining member 58 in the direction of movement of the retaining member 56 and the retaining member 58 (direction of the arrows shown in Figure 2). A culture bag having a bellows structure is a suitable culture container 20 for such a structure. With this, by changing the volume inside the culture container 20 according to the size of the cells to be cultured, the amount of culture medium required for cell culture can be reduced, and consequently the cost of cell culture can be reduced.

[0027] (Modified Version) Figure 3 shows a modified version of the holding structure of the cell sheet laminate 50. In embodiments 1 and 2, the holding member 56 and the first cell sheet 52, and the holding member 58 and the second cell sheet 54 are held in contact with each other. In contrast, in this modified version, porous sheets 57 and 59, to which cells are not attached, are interposed between the holding member 56 and the first cell sheet 52, and between the holding member 58 and the second cell sheet 54, respectively. Both porous sheets 57 and 59 are made of a material that allows culture medium to flow through. As materials for porous sheets 57 and 59, apatite and materials used for the base material of cell sheets can be used. This makes it easier for cells contained in the first cell sheet 52 or cells contained in the second cell sheet 54 to adhere to the porous sheet 57 or 59, and makes it easier to maintain the shape of the cell population obtained in culture.

[0028] Although embodiments of the present invention have been described above, these are merely examples, and various other configurations can be adopted. In the embodiments described above, a circulation pathway for the culture medium is formed, but the culture medium used to culture the cells contained in the first cell sheet 52 and the second cell sheet 54 may be discarded instead of being returned to the culture medium container 30.

[0029] 10 Cell organizing reactor, 20 Culture vessel, 30 Medium container, 40 Control unit, 50 Cell sheet laminate, 52 First cell sheet, 54 Second cell sheet, 60 Medium introduction tube, 62 Pump, 64 Medium discharge tube Cross-reference of related applications

[0030] This application claims priority pursuant to Japanese Patent Application No. 2024-168452, filed with the Japan Patent Office on 27 September 2024, all of which disclosures are incorporated herein by reference in their entirety.

Claims

1. A cell organizing reactor comprising a cell sheet laminate comprising a first cell sheet containing a specific type of cell and a second cell sheet containing a different specific type of cell, which is arranged on at least one side of the first cell sheet, and which supplies a culture medium to the cell sheet laminate.

2. The cell-organizing reactor according to claim 1, wherein the first cell sheet comprises one or more cells selected from the group consisting of muscle-derived cells, skin-derived cells, intestinal-derived cells, heart-derived cells, brain-derived cells, stomach-derived cells, germinal membrane-derived cells, liver-derived cells, kidney-derived cells, and lung-derived cells, and the second cell sheet comprises cells other than those used in the first cell sheet.

3. The cell organizing reactor according to claim 1 or 2, further comprising a holding mechanism including a pair of holding parts for holding the cell sheets in a stacked and sandwiched state, wherein the holding mechanism gradually increases the distance between the pair of holding parts during the cell culture process by supplying culture medium by the culture medium supply part.

4. The cell organizing reactor according to claim 1 or 2, wherein the cell sheet is in a state in which cells are seeded on a porous substrate.

5. The cell organizing reactor according to claim 3, comprising a housing member for housing the cell sheet laminate and the pair of holding parts.

6. The cell organizing reactor according to claim 5, wherein the housing member is expandable or contractible depending on the distance between the pair of holding parts.

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