Cell culture device, cell pharmaceutical preparation, and method for producing cell pharmaceutical preparation

Abstract

One purpose of the present disclosure is to provide a cell culture device from which a string-like aggregate can be removed by a simple operation.　The present embodiment is a cell culture device including a body part having an internal space and a hollow fiber in the internal space. The hollow fiber has a first end having a first end-side opening, and a second end positioned at the end on the opposite side from the first end and having a second end-side opening. The hollow fiber is disposed in the body part such that the first end-side opening opens to the outside of the body part and the second end-side opening opens to the outside of the body part. The lumen of the hollow fiber is partitioned from the internal space in a liquid-tight manner. The body part is configured so as to be capable of supplying a liquid from the outside into the internal space.

Description

Cell culture device, cell therapy preparation, and method for manufacturing cell therapy preparation

[0001] This specification includes the disclosures of Japanese Patent Application No. 2024-173308, which forms the basis for the priority of this application. This disclosure relates to a cell culture device capable of encapsulating cells, a cell therapy preparation in which a transplant material containing cells is placed in the cell culture device, and a method for producing the cell therapy preparation, etc.

[0002] In regenerative medicine, it is necessary to proliferate and culture cells outside the body and prepare a sufficient quantity of high-quality cells. One known technique for proliferating and culturing cells outside the body is to use the lumen of hollow fibers composed of semipermeable membranes (Patent Document 1). It has been reported that even cells that are difficult to proliferate and culture, such as ES cells and iPS cells, can be cultured in large quantities using hollow fibers (Non-Patent Documents 1, 2, and 2). However, previous reports have not investigated methods for supplying cells proliferated and cultured within hollow fibers to clinical settings in a state suitable for transplantation.

[0003] Cell transplantation can be performed using methods such as applying cell suspension, applying cell sheets, or creating and applying organoids. Cell suspension is easy to prepare and allows for transplantation with minimal surgical intervention, but controlling the placement of cells at the transplant site is difficult, and it cannot cover a wide area. On the other hand, cell sheets can cover a wide area at the transplant site, but they are time-consuming to prepare and require invasive surgical procedures involving large incisions. Creating organoids requires advanced skills and time.

[0004] Patent Document 3 reports that in the transplantation of retinal pigment epithelial cells (RPE), the cells can be easily injected under the retina by forming them into string-like aggregates, and a certain area can be covered in the same way as a sheet (Patent Document 3). Patent Document 3 discloses a method for producing string-like aggregates using a device that has a base member having an upper surface provided with one or more grooves.

[0005] Japanese Patent Publication No. 2016-007207, Japanese Patent Publication No. 2018-050498, WO2022 / 230977

[0006] Fujii et al. Cytotechnology. 2020 Apr; 72(2): 227-237Matsushita et al. J Biosci Bioeng. 2019 Oct; 128(4): 480-486

[0007] As described above, Patent Document 3 discloses a method for producing string-like aggregates using a device that includes a base member having an upper surface with one or more grooves. During transplantation, the string-like aggregates produced by this method need to be removed from the grooves of the manufacturing device and transferred to a transport device. However, since string-like aggregates generally have a thin shape with an outer diameter of several hundred micrometers, the operation of removing them from the grooves of the manufacturing device is extremely complicated.

[0008] Therefore, one of the objectives of this disclosure is to provide a cell culture device that can extract string-like aggregates with simple operations.

[0009] Examples of embodiments of the present disclosure are described below. (1) A cell culture device comprising a main body having an internal space and a hollow fiber in the internal space, wherein the hollow fiber has a first end having a first end-side opening and a second end located at the end opposite to the first end and having a second end-side opening, the hollow fiber is arranged in the main body such that the first end-side opening opens to the outside of the main body and the second end-side opening opens to the outside of the main body, the lumen of the hollow fiber is liquid-tightly partitioned from the internal space, and the main body is configured to allow liquid to be supplied to the internal space from the outside. (2) The cell culture device according to (1), wherein the main body includes a torso, the first end-side opening at one end of the torso opens to the outside of the main body, and the second end-side opening at the other end of the torso opens to the outside of the main body. (3) The cell culture device according to any one of (1) to (2), wherein the main body has a mesh structure in at least a part of it. (4) The cell culture device according to any one of (1) to (3), wherein the main body has at least one opening in the main body that communicates with the internal space. (5) The cell culture device according to any one of (1) to (4), wherein the main body has a liquid inlet and a liquid outlet that communicate with the internal space, respectively. (6) The cell culture device according to any one of (1) to (5), wherein the hollow fiber consists of a single hollow fiber. (7) The cell culture device according to any one of (1) to (6), further having a first connecting portion that is detachable from the first connected member, wherein the first end-side opening of the hollow fiber is open to the outside at the first connecting portion. (8) The cell culture device according to any one of (1) to (7), wherein the first connecting portion is connectable to the first connected member such that the inside of the first connected member and the first end-side opening are in spatial communication. (9) The cell culture device according to any one of (1) to (8), wherein the first connecting portion has a female Luer fitting shape. (10) The cell culture device according to any one of (1) to (9), wherein the connecting portion of the first connected member has a male Luer fitting shape corresponding to the female Luer fitting shape of the first connecting portion.(11) The cell culture device according to any one of (1) to (10), wherein the first connected member is a cell extrusion means (e.g., a syringe member). (12) The cell culture device according to any one of (1) to (11), further having a second connecting portion that is detachable from a second connected member in a portion separate from the first connecting portion, and the second end-side opening of the hollow fiber is open to the outside at the second connecting portion. (13) The cell culture device according to any one of (1) to (12), wherein the second connecting portion is connectable to the second connected member such that the inside of the second connected member and the second end-side opening are in spatial communication. (14) The cell culture device according to any one of (1) to (13), wherein the second connecting portion has a male Luer fitting shape. (15) The cell culture device according to any one of (1) to (14), wherein the connecting portion of the second connected member has a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion. (16) The cell culture device according to any one of (1) to (15), wherein the second connected member is a cell injection means (e.g., an injection needle member). (17) The cell culture device according to any one of (1) to (16), further comprising a protective tube made of metal or resin surrounding the outer circumference of the second end of the hollow fiber. (18) The cell culture device according to any one of (1) to (17), wherein the protective tube protrudes to the outside from the second connection portion. (19) The cell culture device according to any one of (1) to (18), wherein the second connection portion has a male Luer fitting shape, the second connected member is an injection needle member including a base having a female Luer fitting shape corresponding to the male Luer fitting shape of the second connection portion and an injection needle disposed at the tip of the base, and the protective tube protrudes to the outside from the second connection portion such that when the second connection portion and the second connected member are connected, the tip of the protective tube and the base end of the injection needle are in contact. (20) The cell culture device according to any one of (1) to (19), wherein the main body is provided with an injection needle in a part separate from the first connection part, and the injection needle is installed such that fluid communication is ensured between the lumen of the injection needle and the second end opening.(21) The cell culture device according to any one of (1) to (20), wherein the injection needle is positioned such that at least a portion of the second end of the hollow fiber enters the lumen of the injection needle. (22) The cell culture device according to any one of (1) to (21), wherein the injection needle has a tapered shape in at least a portion thereof, thereby narrowing the tip of the injection needle. (23) A cell therapy preparation comprising the cell culture device according to any one of (1) to (22) and a transplant material containing cells, wherein the transplant material is positioned in the lumen of the hollow fiber. (24) The cell therapy preparation according to any one of (1) to (23), wherein the cells are at least one selected from stem cells, progenitor cells, somatic cells, and cells differentiated from stem cells or progenitor cells. (25) The cell therapy preparation according to any one of (1) to (24), wherein the cells include retinal pigment epithelial cells. (26) The cell therapy preparation according to any one of (1) to (25), wherein at least a portion of the cells form aggregates in the lumen of the hollow fiber. (27) A method for producing the cell therapy preparation according to any one of (1) to (26), comprising the steps of: placing the cells in the lumen of the hollow fiber; and placing a culture medium in the internal space of the main body and culturing the cells.

[0010] This disclosure provides a cell culture device that allows for the easy removal of string-like aggregates.

[0011] This is a schematic cross-sectional view illustrating an example of the configuration of a cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating an example of the configuration of a cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating an example of the configuration of a cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating the configuration of a hypodermic needle member as an example of a second connectable member connected to the cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating the configuration of a syringe member as an example of a first connectable member connected to the cell culture device according to this embodiment. This is a schematic cross-sectional view showing an example of the configuration near the second connection part of the cell culture device according to this embodiment. This is a schematic cross-sectional view showing an example of the configuration of a hypodermic needle member. This is a schematic cross-sectional view showing the second connection part shown in Figure 5A and the hypodermic needle member shown in Figure 5B connected. This is a schematic cross-sectional view showing an example of the configuration near the second connection part of the cell culture device according to this embodiment, and the second connection part and the hypodermic needle member are connected. This is a schematic cross-sectional view illustrating an example of the configuration of a cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating a configuration in which a cap is attached to the cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating a specific configuration example of the cell culture device according to this embodiment, and is a schematic cross-sectional view taken from a plane along the longitudinal direction of the hollow fiber. This is a schematic cross-sectional view illustrating a specific configuration example of the cell culture device according to this embodiment, and is a schematic side view observed from the first connecting member 102 side. This is a schematic cross-sectional view illustrating a specific configuration example of the cell culture device according to this embodiment, and is a schematic side view observed from the second connecting member 103 side. This is a schematic cross-sectional view illustrating a specific configuration example of the cell culture device according to this embodiment. This is a schematic cross-sectional view illustrating a specific configuration example of the cell culture device according to this embodiment.

[0012] The cell culture device according to this embodiment comprises a main body having an internal space, and a hollow fiber having the internal space, wherein the hollow fiber has a first end having a first end-side opening, and a second end located on the opposite end of the first end and having a second end-side opening, the hollow fiber is arranged in the main body such that the first end-side opening opens to the outside of the main body and the second end-side opening opens to the outside of the main body, the lumen of the hollow fiber is liquid-tightly partitioned from the internal space, and the main body is configured to allow liquid to be supplied to the internal space from the outside.

[0013] This embodiment provides a cell culture device that allows for the easy removal of string-like aggregates. In the cell culture device according to this embodiment, cell aggregates (transplant material) arranged in the lumen of the hollow fibers can be easily removed by injecting a liquid such as physiological saline solution or inserting an extrusion rod through the first end opening or the second end opening. Therefore, by using the cell culture device according to this embodiment, cultured cells can be removed from the cell culture device with simple operations. As a result, complicated tasks such as removing cell aggregates from grooves are not required.

[0014] Furthermore, the cell culture device according to this embodiment allows cells to be cultured in the lumen of hollow fibers composed of a semipermeable membrane and obtained as string-like aggregates. The cultured cells can be transported while contained within the hollow fibers of the cell culture device, and in some cases, the culture vessel used for cultivation can also be transported together. Therefore, it is easy to maintain the quality of the cultured cells, and the range of cell transport is expanded compared to conventional methods.

[0015] Furthermore, the cell culture device according to this embodiment can transport the transplant material to the site of the transplant surgery (e.g., the operating room) while holding the transplant material within the hollow fiber. Also, when removing the transplant material, the cell culture device according to this embodiment may directly load the transplant material coming out of the end opening of the hollow fiber into the cell injection means. Therefore, the cell culture device according to this embodiment does not require the skill of an expert to aspirate cell aggregates from a petri dish or the like and transfer them to the injection needle member for transplantation.

[0016] 1. Cell Culture Device The configuration of the cell culture device according to this embodiment will be described below with reference to the drawings.

[0017] Figure 1 is a schematic cross-sectional view illustrating the configuration of a cell culture device according to this embodiment. In Figure 1, the cell culture device 100 has a main body 11 having an internal space 15. The cell culture device 100 also has a first opening 11a in a part of the main body 11, and a second opening 11b in a part of the main body 11 separate from the first opening 11a. The main body 11 includes, for example, a body portion, a first side wall portion that closes one end of the body portion, and a second side wall portion that closes the end of the body portion opposite to the first side wall portion, with the first opening 11a provided in the first side wall portion and the second opening 11b provided in the second side wall portion. The body portion is not particularly limited, but may be, for example, substantially cylindrical or substantially polygonal prism shape. The hollow fiber 14 has a first end having a first end-side opening 14a, a second end located at the end opposite to the first end, and a second end having a second end-side opening 14b. Furthermore, in this specification, the portion of the hollow fiber 14 located between the first end and the second end is also referred to as the central region. The first end of the hollow fiber 14 is positioned in the main body 11 such that the first end-side opening 14a opens to the outside of the main body 11, specifically positioned in the first opening 11a. Similarly, the second end of the hollow fiber 14 is positioned in the main body 11 such that the second end-side opening 14b opens to the outside of the main body 11, specifically positioned in the second opening 11b. The central region between the first end and the second end of the hollow fiber 14 is positioned to be located in the internal space 15. That is, the hollow fiber 14 is positioned in the main body 11 such that the first end-side opening 14a opens to the outside of the main body 11, the second end-side opening 14b opens to the outside of the main body 11, and the central region is located in the internal space 15. The hollow fiber 14 is installed under tension and held substantially in a straight line. By applying tension, sagging and bending of the hollow fiber can be prevented. The lumen of the hollow fiber 14 is liquid-tightly partitioned from the internal space 15. Furthermore, the main body 11 is configured to allow liquid to be supplied to the internal space 15 from the outside. Note that being configured to allow liquid to be supplied from the outside also means being configured to allow liquid to be discharged to the outside. Also, "opening to the outside" means that the opening is provided so that the inside of the opening is connected to the outside.As will be obvious to those skilled in the art, the opening does not necessarily have to be open to the outside at all times; it may be sealed off from the outside by a cover member, sealing material, or the like as needed.

[0018] One example of a configuration in which the internal space 15 can be supplied with liquid from the outside is a configuration in which the main body has a mesh structure in at least one part. By having at least one part of the main body made of a mesh structure, liquid such as culture medium can be supplied to the internal space 15 through the mesh structure. For example, at least a part of the body may be made of a mesh structure. The mesh density of the mesh structure can be appropriately selected so as to allow for the exchange of culture medium in the internal space 15 without hindering cultivation. Furthermore, it is preferable that the mesh structure has sufficient strength for transport and gripping.

[0019] The main body portion 11 may be configured to have at least one opening. There may be only one opening or there may be multiple openings. If there are multiple openings, it is easier to supply or discharge liquids such as culture medium. The opening may be provided so that it can be opened and closed, for example, by using a cap or the like to select the open or closed state. The opening may have a structure that can be sealed. The opening may be provided in the body portion, for example.

[0020] The main body 11 may have a liquid inlet 107 and a liquid outlet 108 (see Figure 10). For example, the main body 11 may have a first main body opening and a second main body opening, and the first and second main body openings may function as a liquid inlet and a liquid outlet, respectively. The liquid inlet and / or liquid outlet may be provided in the body. The liquid inlet and liquid outlet may be connected through a pipe, and a liquid such as culture medium may be circulated using a pump or the like.

[0021] The lumen of the hollow fiber 14 functions as a space for culturing and holding cells. Cells can be placed in the lumen of the hollow fiber 14 using, for example, a cell extrusion means such as a syringe member. Specifically, cells or a cell suspension can be pushed into the lumen of the hollow fiber 14 from the first end opening 14a or the second end opening 14b of the hollow fiber using a cell extrusion means such as a syringe member containing a cell suspension to be cultured. For example, a needle connected to a syringe member may be inserted into the lumen of the hollow fiber 14, and cells or a cell suspension may be pushed into the lumen of the hollow fiber 14 by pushing the plunger. After filling the hollow fiber with cells to be cultured, both ends of the hollow fiber, specifically the first end opening 14a and the second end opening 14b, may be sealed with a solid material to prevent the cells from moving to other locations. Examples of solid materials include sealants such as resins or stoppers.

[0022] The hollow fiber 14 is not particularly limited, but it is preferable that it consists of a single hollow fiber. That is, in one embodiment, the first end and the second end of the single hollow fiber are positioned at the first opening 11a and the second opening 11b, respectively, so that the lumen of the single hollow fiber opens to the outside of the main body.

[0023] The lumen of the hollow fiber 14 is liquid-tightly partitioned from the internal space 15 of the main body 11, so that even if a liquid such as a culture medium is supplied to the internal space 15, the liquid will not enter the lumen of the hollow fiber 14. In this specification, "liquid-tight" does not refer to the diffusion or passage of components through micropores present on the surface of the hollow fiber, but rather means that a degree of sealing (sealing) is ensured so that liquids such as water do not pass through the gaps in the partitioned space. As described above, the first end of the hollow fiber 14 is positioned in the first opening 11a of the main body 11 such that the first end side opening 14a opens to the outside. In one embodiment, the first end of the hollow fiber 14 is fixed in the first opening 11a with an adhesive such as resin. That is, the cell culture device according to this embodiment may have a potting section that fixes the first end of the hollow fiber 14, which has the first end side opening 14a, to the main body 11, preferably the first opening 11a. Similarly, in one embodiment, the second end of the hollow fiber 14 is fixed to the second opening 11b with an adhesive such as resin so that the second end-side opening 14b opens to the outside. That is, the cell culture device according to this embodiment may have a potting section that fixes the second end of the hollow fiber 14, which has the second end-side opening 14b, to the main body 11, preferably the second opening 11b. In this specification, the statement that the first end-side opening or the second end-side opening opens to the outside of the main body means that the opening can communicate with the space outside the main body. The first end-side opening or the second end-side opening may be opened when injecting cells or when removing cell aggregates, and may be sealed during culture or transport. As a method of arranging the hollow fiber 14 inside the main body 11, for example, both ends of the hollow fiber 14 can be placed in the first opening 11a and the second opening 11b of the main body, respectively, fixed with an adhesive, and then both ends of the hollow fiber can be cut so that the ends of the hollow fiber open to the outside of the main body. Another method involves, for example, positioning both ends of the hollow fiber 14 in the first opening 11a and the second opening 11b, respectively, so that the end openings of the hollow fiber open to the outside of the main body, and then fixing the ends with an adhesive or the like. If the end openings of the hollow fiber are sealed with an adhesive, the ends can be opened by cutting or the like.

[0024] In the cell culture device 100, cell aggregates (transplant material) placed in the lumen of the hollow fibers can be easily removed by injecting a liquid such as physiological saline through the first end opening 14a or the second end opening 14b, or by pushing them out with a plunger or the like. Therefore, by using the cell culture device according to this embodiment, cultured cells can be removed from the cell culture device with simple operations. As a result, complicated work such as removing cell aggregates from grooves is not required. Furthermore, the cell culture device 100 allows cells to be cultured in the lumen of hollow fibers composed of a semipermeable membrane and obtained as string-like aggregates. The cultured cells can be transported while still inside the hollow fibers of the cell culture device, and in some cases, the culture vessel used for culture can be transported together. As a result, it is easy to maintain the quality of the cultured cells, and the range of cell transport is expanded compared to conventional methods. In addition, the cell culture device 100 allows the transplant material to be transported to the site of transplant surgery (e.g., the operating room) while still held inside the hollow fibers. Furthermore, when removing the transplant material, the cell culture device 100 may directly load the transplant material coming out of the end opening of the hollow fiber into the cell injection means. Therefore, the cell culture device 100 eliminates the need for skilled personnel to aspirate cell aggregates from a petri dish or the like and transfer them to the injection needle component for transplantation.

[0025] One aspect of this embodiment is the use of a cell therapy preparation according to this embodiment, which includes the step of removing the implantation material placed in the lumen of the hollow fiber by injecting a liquid from the first end opening or the second end opening or by inserting an extrusion rod.

[0026] Figure 2 is a schematic cross-sectional view illustrating the configuration of the cell culture device according to this embodiment. In Figure 2, the cell culture device 101 further includes a first connecting portion 12 in addition to the basic configuration of the cell culture device 100. The first connecting portion 12 is detachably attached to a first connected member (not shown). Furthermore, the first end-side opening 14a of the hollow fiber opens to the outside at the first connecting portion 12. Specifically, the first end-side opening 14a of the hollow fiber communicates with the outside via the first connecting portion 12. The first connecting portion 12 can be connected to the first connected member such that the inside of the first connected member and the first end-side opening 14a are spatially connected.

[0027] In one embodiment, the first connecting portion 12 has a first connecting portion opening 12a. The first connecting portion opening 12a is spatially in communication with the first end-side opening 14a.

[0028] In one embodiment, the first connecting portion 12 is connectable to the first connected member such that fluid communication is ensured between the inside of the first connected member and the first end-side opening 14a.

[0029] Figure 3 is a schematic cross-sectional view illustrating the configuration of the cell culture device according to this embodiment. In Figure 3, the cell culture device 102 further includes a second connection portion 13 in addition to the basic configuration of the cell culture device 101. The second connection portion 13 is located in a different part from the first connection portion 12 and is detachably connected to a second connected member (not shown). The main body portion 11 includes, for example, a body portion, a first side wall portion that closes one end of the body portion, and a second side wall portion that closes the end of the body portion opposite to the first side wall portion. The first connection portion 12 is provided on the first side wall portion, and the second connection portion 13 is provided on the second side wall portion. The first end-side opening 14a of the hollow fiber 14 opens to the outside at the first connection portion 12, and the second end-side opening 14b of the hollow fiber 14 opens to the outside at the second connection portion 13. Specifically, the first end opening 14a of the hollow fiber 14 is in communication with the outside via the first connecting portion 12, and the second end opening 14b of the hollow fiber 14 is in communication with the outside via the second connecting portion 13. The second connecting portion 13 can be connected to the second connected member such that the inside of the second connected member and the second end opening 14b are in spatial communication.

[0030] In one embodiment, the second connection portion 13 has a second connection portion opening 13a. The second connection portion opening 13a is spatially in communication with the second end-side opening 14b.

[0031] In one embodiment, the second connecting portion 13 is connectable to the second connected member such that fluid communication is ensured between the inside of the second connected member and the second end-side opening 14b.

[0032] The first connecting portion 12 and the second connecting portion 13 may be integrally formed from the same material as the main body portion 11, or they may be formed as separate components from the same or different materials as the main body portion 11. If the connecting portions are formed as separate components from the main body portion, they can be integrated by, for example, heat fusion, but are not limited to this.

[0033] The first connecting portion 12 is configured to be detachably attached to an external first connected member (not shown). The second connecting portion 13 is configured to be detachably attached to an external second connected member (not shown). For example, the first connecting portion 12 can be connected to the first connected member so as to ensure fluid communication, and the second connecting portion 13 can be connected to the second connected member so as to ensure fluid communication. The cell culture device according to this embodiment has hollow fibers 14 made of a semipermeable membrane in the internal space 15 of the main body portion 11, and the hollow fibers 14 have a first end-side opening 14a and a second end-side opening 14b opposite to the first end-side opening 14a. The hollow fibers 14 are arranged in the main body portion 11 such that the first end-side opening 14a opens at the first connecting portion 12. The hollow fibers 14 are also arranged in the main body portion 11 such that the second end-side opening 14b opens at the second connecting portion 13. For example, the hollow fiber 14 is positioned in the internal space 15 between the first connection part 12 and the second connection part 13 such that the first end opening 14a communicates with the first connection part opening 12a and the second end opening 14b communicates with the second connection part opening 13a. The first connection part opening 12a and the second connection part opening 13a are liquid-tightly separated from the internal space 15 of the main body part 11. The hollow fiber 14 may also be fixed by a potting part, which can fix the hollow fiber 14 to the main body part 11 such that the first end opening 14a and the second end opening 14b of the hollow fiber 14 open to the outside at the first connection part 12 and the second connection part 13, respectively. For example, the potting part can be provided at the first opening 11a and the second opening 11b of the main body part 11, and in one embodiment, the potting part can ensure liquid tightness of the first connection part opening 12a and the second connection part opening 13a to the internal space 15. The potting area can be formed using a resin such as an adhesive.

[0034] The first connection opening 12a and the second connection opening 13a do not necessarily need to be in constant communication with the outside, and may be sealed off from the outside by a lid member or sealing material as needed. For example, the first connection opening 12a may be sealed off from the outside by a lid member. Specifically, during cell culture and transport, it is sealed off from the outside by a lid member or sealing material. Also, for example, the first connection opening 12a may be filled with a resin material such as potting resin. That is, even if the first connection opening exists in the first connection part during the manufacturing process of the cell culture device, in the final product, the cell culture device, the opening may be filled with another material such as potting resin. Such lid members and sealing materials can be removed as appropriate depending on the use of the cell culture device. Specifically, the lid members and sealing materials are removed when placing cells in the lumen of the hollow fiber or when administering cells to the affected area. The same applies to the second connection opening 13a.

[0035] In one embodiment, the first connection portion 12 can be connected to a first connected member, which may be an external element of the cell culture device according to this embodiment, so as to ensure fluid communication, and the second connection portion 13 can be connected to a second connected member, which may be an external element of the cell culture device according to this embodiment, so as to ensure fluid communication.

[0036] The shapes of the first connecting portion 12 and the second connecting portion 13 are independent of each other, but it is preferable that the first connecting portion 12 and the second connecting portion 13 have a shape that allows them to be connected by a fitting, which is a common optional connection method. The first connecting portion 12 and the second connecting portion 13 are formed, for example, in the shape of a lure fitting as defined by the International Standard (ISO), for example, in the shape of a male lure fitting or a female lure fitting.

[0037] Examples of the first connected member or the second connected member include, for example, cell injection means for arranging cells at the transplantation site, cell extrusion means for extruding cells in the lumen of the hollow fiber, and the like. In one embodiment, the first connected member connected to the first connection portion 12 is cell extrusion means for extruding cells in the lumen of the hollow fiber. Also, in one embodiment, the second connected member connected to the second connection portion 13 is cell injection means for arranging cells at the transplantation site.

[0038] The cell extrusion means is not particularly limited, and examples thereof include a syringe member. The cell extrusion means can be configured as an external element not included in the cell culture device according to the present embodiment. By the cell extrusion means, the contents present in the lumen of the hollow fiber can be sent out to the outside. For example, a syringe member filled with a liquid is connected to the first connection portion, and the liquid in the syringe member is injected into the lumen of the hollow fiber from the opening on the first end side by an extrusion operation, whereby the contents present in the lumen of the hollow fiber can be moved to the opening on the second end side. Also, examples of the extrusion rod include a plunger with a tip covered with a PTFE or rubber material, etc. By inserting the extrusion rod into the lumen of the hollow fiber from the opening on the first end side, the contents can be extruded to the opening on the second end side. The plunger as the extrusion rod can also be used in a set, that is, in combination with the syringe member. That is, the plunger can be arranged inside the syringe member, and when an extrusion operation is performed, the tip of the plunger protrudes from the tip of the syringe member, and the contents present in the lumen of the hollow fiber can be configured to be extruded by the tip of the plunger. As such cell extrusion means, for example, a microsyringe is preferably mentioned. Regarding the plunger of the microsyringe, by appropriately selecting the diameter of its needle according to the inner diameter of the hollow fiber and its length according to the length of the hollow fiber, the contents in the hollow fiber can be efficiently extruded. Commercially available products may be used for the cell extrusion means. In other words, the connection portion of the first connection portion can be designed so that commercially available products can be used as the cell extrusion means.

[0039] Therefore, by providing the first connecting portion 12, cell aggregates inside the hollow fiber can be easily removed.

[0040] The cell injection means is not particularly limited, but examples include injection needle components. The cell injection means may be configured as an external element not included in the cell culture device according to this embodiment. The injection needle component has an injection needle, which includes a transplant needle or transplant cannula. By inserting the injection needle (e.g., transplant needle or transplant cannula) into the tissue from the tip of the needle, the tip of the needle can reach the site to transplant cells, and cells can be placed at the transplant site by pushing them out from the hollow fiber lumen. The material of the transplant needle or transplant cannula is not particularly limited, but for example, it may be resin or metal, and metal is preferred. The cell culture device according to this embodiment can be used for transplantation and can therefore also be understood as a transplantation device. The second connected member is not limited to a cell injection means for placing cells at the transplant site, but may also be an injection needle component that is not a cell injection means. That is, the injection needle component connected as the second connected member does not have to be used as a cell injection means. In this case, cell aggregates can be transferred from the injection needle component to a cell injection means such as a transplant needle or transplant cannula. A commercially available cell injection device may be used. In other words, the connection portion of the second connection can be designed so that a commercially available cell injection device can be used.

[0041] Therefore, by providing the second connection part 13, the cell culture device can be used for transplantation. As a result, the complicated operation of transferring cell aggregates to the transplantation needle (transplantation injection needle component) immediately before transplantation is eliminated, making it possible to handle an increase in the number of transplantation cases. Furthermore, even if not used for transplantation, by providing the second connection part 13, cell aggregates can be easily transferred from the injection needle component to a cell injection means such as a transplantation needle or transplantation cannula, thus simplifying the operation of transferring cell aggregates to the transplantation needle (transplantation injection needle component).

[0042] In one embodiment, the first connecting portion 12 has a female Luer fitting shape. The connecting portion of the first connected member has a male Luer fitting shape corresponding to the female Luer fitting shape of the first connecting portion.

[0043] In one embodiment, the second connecting portion 13 has a male Luer fitting shape. The connecting portion of the second connected member has a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion 13.

[0044] In one embodiment, the first connecting portion 12 has a female Luer fitting shape, and the second connecting portion 13 has a male Luer fitting shape. The connecting portion of the first connected member has a male Luer fitting shape corresponding to the female Luer fitting of the first connecting portion 12, and the connecting portion of the second connected member has a female Luer fitting shape corresponding to the male Luer fitting of the second connecting portion 13. In this embodiment, when the first connecting portion 12 has a female Luer fitting shape, it is preferable that the second connecting portion 13 has a male Luer fitting shape, and when the first connecting portion 12 has a male Luer fitting shape, it is preferable that the second connecting portion 13 has a female Luer fitting shape. However, the present invention is not particularly limited to these forms.

[0045] In one embodiment, the first connected member is a syringe member and the second connected member is an injection needle member.

[0046] Figure 4A is a schematic cross-sectional view showing the configuration of the injection needle member, and Figure 4B is a schematic cross-sectional view showing the configuration of the syringe member. In Figure 4A, the injection needle member 30 includes a base 31 and an injection needle 32 positioned at the tip of the base 31, and the base 31 has a female Luer fitting shape corresponding to a second connecting portion 13 having a male Luer fitting shape. In one embodiment, the second connected member is an injection needle member including a base 31 having a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion 13, and an injection needle 32 positioned at the tip of the base 31. The injection needle may have a tapered shape in at least a portion of it. By having a tapered shape in the injection needle, an injection needle can be used in which the tip is thinner than the base end. In Figure 4B, the syringe member 40 includes a tubular body 41 and a tip portion 42 positioned at the tip of the tubular body, and may also include a plunger 43. The plunger may be configured such that when an extrusion operation is performed, the plunger tip protrudes from the tip of the syringe member, and the contents present in the lumen of the hollow fiber are pushed out by the plunger tip. The tip portion 42 has a male Luer fitting shape that corresponds to the first connecting portion 12 which has a female Luer fitting shape.

[0047] In one embodiment, the Luer interface is threaded (in this case, the configuration is called a “Luer lock” configuration), and the two sides can be connected by relative rotation, which may be combined with a compressive load. In other words, in one embodiment of a Luer lock, rotation, sometimes together with compression, can be used to engage the threads of a male Luer fitting configured to engage with the flange of a female Luer fitting, thereby connecting the members and ensuring fluid communication between them. That is, by engaging the threads of a male Luer fitting configured to engage with the flange of a female Luer fitting by rotation, the members can be securely connected and fluid communication between them can be ensured. In one embodiment, a tapered interface shape can be used to provide a Luer engagement using compression without threading or rotation (such a configuration may be called a “slip-on” or “conical” Luer configuration).

[0048] Figure 5A is a schematic cross-sectional view showing the configuration near the second connection portion of the cell culture device according to this embodiment. In Figure 5A, a second connection portion 13 having a male Luer fitting shape is arranged at the end of the body portion constituting the main body portion 11. In Figure 5A, the second connection portion 13 is composed of a second connection portion component, which is a separate member from the main body portion 11. The second connection portion component consists of a cylindrical portion arranged inside the main body portion 11 and a connection portion arranged outside the main body portion 11 and connectable to the second connected member. The cylindrical portion of the second connection portion component is inserted into the main body portion through an opening (second opening) provided in the main body portion and is arranged inside the main body portion 11. The second connection portion component has an internal opening of the cylindrical portion inside the cylindrical portion and an internal opening of the connection portion inside the connection portion, and the internal opening of the cylindrical portion and the internal opening of the connection portion are in communication, constituting the second connection portion opening 13a. The second end of the hollow fiber 14 is arranged inside the second connection portion opening 13a. The second end opening 14b of the hollow fiber 14 opens to the outside at the end of the second connecting component. Potting resin 18 is sealed in the second connecting opening 13a of the second connecting component, and the potting resin 18 fixes the second end of the hollow fiber 14 to the second connecting component, ensuring liquid-tightness between the internal space of the main body and the outside. Furthermore, the hollow fiber 14 is positioned to pass through the center of the second connecting opening 13a.

[0049] Figure 5B is a schematic cross-sectional view showing the configuration of the injection needle member 30, wherein the base portion 31 of the injection needle member 30 (specifically, the inner wall of the base portion) has a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion 13 shown in Figure 5A.

[0050] Figure 5C is a schematic cross-sectional view showing the state in which the second connecting portion 13 shown in Figure 5A and the injection needle member 30 shown in Figure 5B are connected. By connecting the second connecting portion and the injection needle member (second connected member), it becomes possible to inject the cell aggregates present inside the hollow fiber into the target tissue through the injection needle 32.

[0051] Figure 6 is a schematic cross-sectional view showing an example of the configuration near the second connection portion of the cell culture device according to this embodiment, and is a schematic cross-sectional view showing the state in which the second connection portion and the injection needle member are connected. The configuration near the second connection portion shown in Figure 6 includes, in addition to the configuration shown in Figure 5A or 5C, a protective tube 17 surrounding the outer circumference of the second end of the hollow fiber 14.

[0052] The protective tube 17 may be made of metal or resin. Furthermore, the protective tube 17 may be formed from a single layer or from two or more layers. In Figure 6, the protective tube 17 is formed from two layers: a first protective tube 17a and a second protective tube 17b. For example, the first protective tube 17a is made of metal, and the second protective tube 17b is made of resin. The spaces between the hollow fiber and the protective tube, between the protective tubes themselves, and / or between the protective tube and the second connecting component (the openings) may be sealed with adhesive.

[0053] In Figure 6, the second end of the hollow fiber, the first protective tube 17a, and the second protective tube 17b protrude from the opening end of the opening of the second connecting component. That is, the protective tubes protrude to the outside from the second connecting component. Preferably, the protective tube 17 protrudes to the outside from the second connecting component such that when the second connecting component and the second connected component (injection needle component) are connected, the tip of the protective tube and the proximal end of the injection needle (the end closer to the hollow fiber) come into contact. This reduces or eliminates the gap that may occur between the opening on the second end of the hollow fiber and the proximal end of the injection needle (the end closer to the hollow fiber) when the second connecting component and the second connected component (injection needle component) are connected. This ensures efficient fluid communication between the lumen of the injection needle and the opening on the second end.

[0054] Figure 7 is a schematic cross-sectional view illustrating the configuration of the cell culture device 103 according to this embodiment. In Figure 7, the cell culture device 103 includes, in addition to the basic configuration of the cell culture device 101, an injection needle 16 located in a part of the main body 11 separate from the first connection portion 12. The injection needle 16 is a component of the cell culture device. The injection needle 16 may be configured to be detachable or non-detachable. The injection needle 16 is installed so as to ensure fluid communication between the lumen of the injection needle and the second end-side opening 14b. The main body 11 includes, for example, a body portion, a first side wall portion that closes one end of the body portion, and a second side wall portion that closes the end of the body portion opposite to the first side wall portion. The first connection portion 12 is provided on the first side wall portion, and the injection needle 16 is provided on the second side wall portion. The first end-side opening 14a of the hollow fiber 14 communicates with the lumen of the injection needle 16 and may be open at the first connection portion 12. The second end opening 14b of the hollow fiber 14 is positioned to ensure fluid communication with the lumen of the injection needle 16. The injection needle may have a tapered shape in at least part of it. By having a tapered shape, it is possible to use an injection needle whose tip is thinner than the end on the hollow fiber side.

[0055] The injection needle includes a transplant needle or transplant cannula. By inserting the injection needle (e.g., a transplant needle or transplant cannula) into the tissue from the tip, the tip of the needle can reach the site where cells are to be transplanted, and the cells can be placed at the transplant site by pushing them out from the hollow fiber lumen. The material of the transplant needle or transplant cannula is not particularly limited, but may be resin or metal, for example, and metal is preferred. Since the cell culture device according to this embodiment can be used for transplantation, it can also be understood as a transplantation device or an administration device. Note that this embodiment is not limited to a cell injection means.

[0056] In the cell culture device 103 according to this embodiment, cell aggregates (transplant material) arranged in the lumen of the hollow fiber can be easily removed from the injection needle by injecting a liquid such as physiological saline solution through the first end opening or by inserting an extrusion rod. Alternatively, in the cell culture device 103 according to this embodiment, the injection needle can be inserted into the tissue from the tip of the needle to reach the site where cells are to be transplanted, and the cells can be pushed out from the lumen of the hollow fiber to position them at the transplantation site.

[0057] In one embodiment, the end of the second end of the hollow fiber 14 may be inserted into the lumen of the proximal end of the injection needle. In Figure 7, the end of the second end of the hollow fiber 14 is inserted into the lumen of the proximal end of the injection needle, so that at least a portion of the second end of the hollow fiber 14 enters the lumen of the injection needle. By adopting such a structure, efficient fluid communication can be ensured between the lumen of the injection needle and the opening on the second end side.

[0058] Furthermore, in one embodiment, in a configuration in which the end of the second end of the hollow fiber 14 is inserted into the lumen of the injection needle, the inner wall of the proximal end of the injection needle can be stepped to create a stepped shape. The diameter of the lumen of the injection needle in the portion with this stepped shape can be made to substantially match the outer diameter of the inserted hollow fiber. By adopting such a structure, the step difference between the inner wall of the hollow fiber and the inner wall of the injection needle at the connection portion between the inner wall of the injection needle and the opening on the second end side of the hollow fiber can be reduced or eliminated. This makes it possible to efficiently transfer cell aggregates within the hollow fiber into the lumen of the injection needle.

[0059] As shown in Figure 8, the cell culture device according to this embodiment may have a first cap 21 that is detachable from the first connection part 12. The cell culture device according to this embodiment may also have a second cap 22 that is detachable from the second connection part 13. By having the first cap 21 and the second cap 22, liquid-tight or airtight seals can be achieved between the lumen and the outside of the hollow fiber 14. This allows the cell culture device to be immersed in the culture medium, either entirely or partially, during cell culture, thereby supplying the culture medium to the internal space 15 while ensuring liquid-tight or airtight seals between the lumen and the outside of the hollow fiber 14.

[0060] In one embodiment, the first cap 21 may have a first protrusion 21a that fits into the first connection opening 12a when attached to the first connection 12. The second cap 22 may have a second protrusion 22a that fits into the second connection opening 13a when attached to the second connection 13. By having the first protrusion 21a that fits into the first connection opening 12a of the first cap 21, or by having the second protrusion 22a that fits into the second connection opening 13a of the second cap 22, the first end-side opening 14a or the second end-side opening 14b of the hollow fiber 14 can be sealed, effectively suppressing leakage or detachment of cells from the first end-side opening 14a or the second end-side opening 14b. The first end-side opening 14a or the second end-side opening 14b of the hollow fiber 14 may be sealed using a sealant such as resin. Furthermore, the first end opening 14a or the second end opening 14b of the hollow fiber 14 does not need to be sealed. In particular, even if it is not sealed with a sealant, due to surface tension, etc., leakage or detachment of cells from the first end opening 14a or the second end opening 14b of the hollow fiber is minimal.

[0061] The semipermeable membrane that makes up hollow fibers is a membrane that does not allow cells to pass through, but allows substances necessary for cell culture and proliferation to pass into the lumen of the hollow fiber, while allowing unwanted waste products to pass out to the outside of the microtubule. The material for such a semipermeable membrane is not particularly limited, but examples include any or a combination selected from the group consisting of polylactic acid (PLA), polyglycolic acid (PGA), lactic acid-glycolic acid copolymer (PLGA), polyhydroxy acid, polycaprolactone, polycarbonate, polyamide, polyethylene, polyurethane, polyarylate, polysulfone, polyethersulfone, polyester, polystyrene, polyvinyl alcohol, polyvinyl acetate, polyvinyl chloride, polyvinyl fluoride, polyvinylimidazole, chlorosulfonated polyolefin, polyethylene oxide, polyphosphazene, polyamino acids, polyorthoesters, polyacetal, polycyanoacrylate, polytetrafluoroethylene (PTFE), biodegradable polyurethane, polyvinylidene fluoride, polytetrafluoroethylene, cellulose acetate, polyacrylonitrile polyacrylate, ethylene-vinyl acetate polymer, acyl-substituted cellulose acetate, polymethyl methacrylate, polypropylene, and regenerated cellulose, as well as their derivatives.

[0062] The pore size of the pores on the surface of the hollow fiber is preferably 0.01 μm to 20 μm, preferably 0.05 μm to 10 μm, and preferably 0.10 μm to 5 μm. By adjusting the pore size of the hollow fiber, the permeability of substances into and out of the hollow fiber can be controlled, thereby improving cell proliferation and maintenance.

[0063] The thickness of the hollow fiber is preferably 10 μm to 100 μm, and more preferably 10 μm to 50 μm. By adjusting the thickness of the hollow fiber, it is possible to control the permeability of substances inside and outside the hollow fiber, as well as control the shape retention (strength) of the hollow fiber.

[0064] The inner diameter of the hollow fiber is not particularly limited as long as it contains the cells necessary for transplantation and allows them to proliferate. The inner diameter of the hollow fiber is, for example, 20 μm to 2000 μm, preferably 50 μm to 1000 μm, preferably 100 μm to 500 μm, preferably 130 μm to 300 μm, and preferably 150 μm to 270 μm.

[0065] The length of the hollow fiber is not particularly limited, as long as it contains the cells necessary for transplantation and allows them to proliferate. The hollow fiber has sufficient length so that the end openings at both ends communicate with the first connection opening 12a of the first connection part 12 and the second connection opening 13a of the second connection part 13.

[0066] The inner diameter and length of the hollow fiber are appropriately optimized within the range described above, depending on the purpose of transplantation and the site of administration, and are not particularly limited. For example, in ophthalmic surgery, the outer diameter of the injection needle used is, for example, 23G (0.65 mm) or 25G (0.5 mm), and the standard (non-thin-walled) inner diameter in that case is, for example, 0.26 mm (@25G) or 0.34 mm (@23G). Furthermore, from the viewpoint of invasiveness, the outer diameter of the cannula tip is preferably 0.3 mm or less. In that case, the outer diameter of the hollow fiber is preferably 300 μm or less, and considering the thickness of the semipermeable membrane (e.g., 10 μm to 50 μm), the inner diameter of the hollow fiber is preferably about 50 μm to 280 μm.

[0067] The cells to be cultured are supplied through the end openings of the hollow fiber. The cells may be supplied through either the first end opening 14a or the second end opening 14b of the hollow fiber. In one embodiment, after the cells are placed in the lumen of the hollow fiber, the first connection opening 12a and / or the second connection opening 12b may be fitted with water-resistant and / or air-blocking caps (first cap or second cap) (see Figure 8).

[0068] The "cells" enclosed within the hollow fibers are not particularly limited and may be at least one selected from stem cells, progenitor cells, somatic cells, and cells differentiated from stem cells or progenitor cells. The cells may be stem cells, progenitor cells, or somatic cells. Examples of stem cells and progenitor cells include pluripotent stem cells such as embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells); hematopoietic stem cells and progenitor cells, neural stem cells and progenitor cells, hepatic stem cells and progenitor cells, pancreatic stem cells and progenitor cells, skin stem cells and progenitor cells, osteochondral stem cells and progenitor cells, and mesenchymal stem cells including adipose stem cells, cardiomyocyte stem cells, and dental pulp stem cells. Examples of somatic cells include differentiated cells such as lymphocytes, epithelial cells, endothelial cells, muscle cells, fibroblasts (skin cells, etc.), hair cells, hepatocytes, gastric mucosal cells, intestinal cells, spleen cells, pancreatic cells (exocrine pancreatic cells, etc.), brain cells, lung cells, kidney cells, eye-related cells, adipocytes, and glial cells. Differentiated cells may also be derived from pluripotent stem cells or stem cell / progenitor cells.

[0069] ES cells may be any established ES cell lines. Examples of ES cell lines that can be used include, but are not limited to, the clinical human ES cell lines provided by the Human ES Cell Research Center, Institute of Medical and Biological Sciences, Kyoto University, and the human ES cell lines provided by RIKEN BRC (KhES-1, KhES-1_Crx::Venus, KhES-1_Rx::Venus).

[0070] As for iPS cells, not only iPS cells derived from the patient's own somatic cells but also various iPS cell lines can be used. As for iPS cell lines, various iPS stock cell lines provided by the Kyoto University iPS Cell Research Foundation, healthy donor-derived iPS cell lines provided by RIKEN BRC, KVs09, CLs23, etc. can be used, but are not limited to these.

[0071] The species of cells is not particularly limited and can be appropriately selected depending on the purpose. For human transplantation, primate-derived cells, especially those derived from monkeys or humans, are preferred, and human-derived cells are more preferred.

[0072] The cell culture device according to this embodiment is particularly suitable for administering cells to a transplantation site using cell injection means such as injection needles or cannulas. Examples of such cells include ophthalmic cells such as corneal epithelial cells, retinal pigment epithelial cells, neuroretinal cells, conjunctival epithelial cells, limbal epithelial cells, corneal endothelial cells, corneal stromal cells, iris stromal cells, scleral cells, iris pigment epithelial cells, ciliary epithelial cells, optic nerve cells, subliminal fibroblasts, subconjunctival fibroblasts, lacrimal gland cells, meibomian gland cells, goblet cells, lenticular epithelial cells, and eyelid epithelial cells. Preferred ophthalmic cells are retinal pigment epithelial cells, neuroretinal cells, and lacrimal gland cells, with retinal pigment epithelial cells being the most preferred. In addition to ophthalmic cells, pancreatic β-cells, hepatic cells, neural glial cells, and neural progenitor cells are also suitable for transplantation using the cell culture device according to this embodiment.

[0073] In the cell culture device according to this embodiment, the main body is preferably made of a transparent material, and preferably a colorless and transparent material. Also, in the cell culture device according to this embodiment, the hollow fibers are preferably made of a transparent material, and preferably a colorless and transparent material. "Transparent material" refers to a material that transmits visible light, regardless of whether it modifies the true color of the contents. For example, an amber-colored material allows the contents to be seen through it, and when viewed in this way, it appears amber-colored, but this also falls under the definition of transparent as defined here. "Colorless and transparent" means that it is both transparent and colorless. This is not an absolute term, as articles that are absolutely transparent or colorless are rare, if any. This term is used in the pharmaceutical industry, for example, to refer to a material that appears colorless and transparent.

[0074] It is preferable that hollow fibers be made of a material that appears transparent when wetted with a liquid, making it easier to confirm the presence of cells. By using hollow fibers made of a material that becomes transparent when wetted with water, quality control of transplant materials can be performed non-destructively and easily. As an example of such a material, our experiments have shown that some PVDF membranes used for virus removal are opaque when dry, but the hollow fibers become transparent when wetted with water. In addition, our experiments have shown that some thin PVDF membranes, regenerated cellulose membranes, and PMMA membranes are opaque when dry, but the hollow fibers become transparent when wetted with water.

[0075] Furthermore, one embodiment of this embodiment may be a cell culture device kit comprising the cell culture device according to this embodiment and the first connected member and / or second connected member. In one embodiment, the first connected member is a cell extrusion means (e.g., a syringe member), and the second connected member is a cell injection means (e.g., an injection needle member).

[0076] 2. Cell Therapeutic Preparation The cell therapy preparation according to this embodiment includes a cell culture device according to this embodiment, and a transplant material containing cells is arranged in the lumen of a hollow fiber. In other words, the cell therapy preparation according to this embodiment includes a cell culture device according to this embodiment and a transplant material containing cells, and the transplant material is arranged in the lumen of a hollow fiber.

[0077] After placing cells in the lumen of the hollow fibers of the cell culture device according to this embodiment, a culture medium can be supplied to the internal space of the main body, and the cells can be grown under appropriate culture conditions. Specifically, as described above, after placing cells in the lumen of the hollow fibers of the cell culture device according to this embodiment, a culture medium can be supplied to the internal space of the main body, and the cells can be grown under appropriate culture conditions. This makes it possible to produce a cell therapy preparation in which a transplant material containing cells is placed in the lumen of the hollow fibers of the cell culture device according to this embodiment. The cells that can be used are as described above.

[0078] Within the lumen of the hollow fiber, at least a portion of the cells form aggregates. In the case of adhesive cells, cells cultured and grown within hollow fibers composed of a semipermeable membrane form aggregates. For example, in the case of retinal pigment epithelial cells, the aggregates formed are similar to the string-like aggregates produced by the device described in WO2022 / 230977, but may also include smaller fragments. The inventors have confirmed that such fragments, like string-like aggregates, may have high efficacy as implantation materials.

[0079] The cell therapy formulation according to this embodiment may, in addition to the cell culture device, include a pharmacologically acceptable carrier or medium, specifically, sterile water, physiological saline, culture medium, physiological buffer such as PBS, preservatives, surfactants, stabilizers, excipients, antiseptics, binders, reducing agents, or isotonic agents. These may be used individually or in combination of two or more. If necessary, a cryopreservative may be added, the formulation may be frozen and stored, and thawed before use.

[0080] The method for producing a cell-based pharmaceutical preparation according to this embodiment includes the steps of placing cells in the lumen of a hollow fiber and placing a culture medium in the internal space of the main body and culturing the cells.

[0081] The lumen of the hollow fiber functions as a space for culturing and holding cells, and the cells to be cultured can be placed in the lumen of the hollow fiber using, for example, a cell extrusion means such as a syringe member. Specifically, cells or cell suspensions can be pushed into the lumen of the hollow fiber from the first end opening or the second end opening of the hollow fiber using a cell extrusion means such as a syringe member containing a cell suspension housing. For example, a needle connected to a syringe member may be inserted into the lumen of the hollow fiber, and cells or cell suspensions may be pushed into the lumen of the hollow fiber by pushing in a plunger. After filling the hollow fiber with cells to be cultured, both ends of the hollow fiber may be sealed with a solid material to prevent the cells from moving to other locations. Examples of solid materials include sealants such as resins or stoppers.

[0082] The internal space 15 of the main body 11 is configured to allow liquid to be supplied from the outside. For example, the main body is configured to have a mesh structure in at least one part, or the main body is configured to have one or more openings. This allows liquid such as culture medium to be supplied or discharged.

[0083] In the culture process, it is preferable that the culture medium is supplied to the internal space 15 such that at least the entire hollow fiber is immersed in the culture medium. For example, the cell culture device may be positioned so that its entire body is immersed in the culture medium. Alternatively, the culture process may be carried out while circulating the culture medium in the internal space 15 using a liquid inlet and liquid outlet provided in a part of the main body.

[0084] The culture apparatus may have known means necessary for cell culture, such as means for adjusting the culture temperature, means for adjusting the oxygen and carbon dioxide concentrations, means for exchanging the culture medium, and means for supplying additional components.

[0085] The culture medium and culture conditions are determined appropriately depending on the cells. For example, the cell density within the hollow fiber is 5 × 10⁻⁶. ７ The cell density is 5 × 10⁻¹⁴ cells / mL or higher, preferably 5 × 10⁻¹⁴ cells / mL or higher. ７ cells / mL ~ 1 x 10 ８ The concentration is cells / mL, preferably 7.5 × 10⁶. ７ cells / mL ~ 1 x 10 ８ The value is cells / mL.

[0086] During cell culture, it is preferable that the first and second connection points of the cell culture device be sealed with caps or the like. This helps to maintain the quality of the transplant material.

[0087] After the cell processing is complete, the cell culture device may be washed as needed, including the outer and inner walls of the main body and the outer circumference of the hollow fibers. Alternatively, the device can be transported to the next process while retaining the culture medium within its internal space. For example, the cell culture device, immersed in the culture medium, can be transported to the next process along with the culture vessel. Therefore, cell therapy preparations can be provided to medical institutions with the transplantation material already filled, without freezing. The opening may be closed or left open.

[0088] As mentioned above, cell therapy preparations can be transported with the cell culture device placed in the culture vessel. Furthermore, if transported under low-oxygen conditions (especially anaerobic conditions), the quality of the cells encapsulated within the tubular structure (hollow fiber) can be maintained for approximately 30 days.

[0089] The cell therapy preparation according to this embodiment may be stored or transported in a frozen state. Freezing the cell therapy preparation (or transplant material) can extend its shelf life and protect it from the effects of transportation.

[0090] As a preferred example of a cell therapy preparation according to this embodiment, a cell therapy preparation containing retinal pigment epithelial cells is described below.

[0091] Retinal pigment epithelial (RPE) cells refer to the epithelial cells that make up the retinal pigment epithelium, and their progenitor cells. RPE cells may be isolated from the patient or induced from stem cells such as pluripotent stem cells. Since the number of RPE cells that can be obtained from a patient is limited, it is necessary to proliferate and expand the RPE cells for transplantation. In this embodiment, a cell therapy preparation can be obtained in which RPE cells are pre-filled in a cell culture device by culturing the cells in hollow fibers made of a semipermeable membrane.

[0092] The culture method for RPE cells basically follows WO2022 / 230977. As the basal medium, any medium commonly used for culturing animal cells can be used. For example, as the basal medium, BME medium, BGJb medium, CMRL 1066 medium, Glasgow MEM (GMEM) medium, Improved MEM Zinc Option medium, IMDM medium, Medium 199 medium, Eagle MEM medium, αMEM medium, DMEM medium, F-12 medium, DMEM / F12 medium, IMDM / F12 medium, Ham medium, RPMI 1640 medium, Fischer's medium, or a mixture of these can be used.

[0093] The culture medium may be serum-containing or serum-free. Serum-free medium may contain a serum substitute. The serum substitute may be a commercially available product, such as Knockout™ Serum Replacement (KSR), Chemically-defined Lipid concentrated (manufactured by Life Technologies), or Glutamax. ＴＭ You can use the following supplements: (manufactured by Life Technologies), B27 (manufactured by Life Technologies), N2 supplement (manufactured by Life Technologies), and ITS supplement (manufactured by Life Technologies).

[0094] The serum-free culture medium may optionally contain fatty acids or lipids, amino acids (e.g., non-essential amino acids), vitamins, growth factors, cytokines, antioxidants, 2-mercaptoethanol, pyruvate, buffers, inorganic salts, etc.

[0095] A ROCK inhibitor may be added to the medium. Examples of the ROCK inhibitor include, for example, Y-27632 dihydrochloride, Y-27632, Fasudil Hydrochloride, Chroman 1, SLx-2119, HSD1590, GSK269962A hydrochloride, Exoenzyme C3, clostridium botulinum, Ripasudil, Afuresertib, Thiazovivin, GSK269962A, RKI-1447, Y-33075, GSK429286A, AT13148, H-1152 dihydrochloride, Y-33075 dihydrochloride, LX7101, SAR407899, ROCK-IN-2, Afuresertib hydrochloride, Hydroxyfasudil, GSK180736A, BDP5290, SR-3677, CCG-222740, CMPD101, Rho-Kinase-IN-1, SAR407899 hydrochloride, ROCK inhibitor-2, ZINC00881524, H-1152, Hydroxyfasudil hydrochloride, Fasudil, ROCK2-IN-2, Verosudil, SB-772077B dihydrochloride, GSK-25, CRT0066854 hydrochloride, Ripasudil free base, ROCK-IN-1, etc. Preferably, they are Y-27632 dihydrochloride and Y-27632. The concentration of the ROCK inhibitor contained in the medium is usually 0 μM to 20 μM, preferably 2 μM to 10 μM. Note that it is preferable to add the ROCK inhibitor when aggregating RPE cells, but the ROCK inhibitor is not essential for the preparation after culture.

[0096] The density of the RPE cells to be cultured is not particularly limited, but is usually 2.5×10 ３ cells / mL or more, preferably 2.5×10 ３ cells / mL to 5×10 ５ cells / mL, more preferably 1×10 ５ cells / mL to 2×10 ５ cells / mL.

[0097] The culture time for RPE cells is not particularly limited, but is usually 1 to 30 days, preferably 2 to 7 days.

[0098] The culture temperature is, for example, about 30°C to about 40°C, preferably about 37°C. ２ The concentration is, for example, about 1% to about 10%, preferably about 5%.

[0099] RPE cells cultured within hollow fibers form aggregates resembling string-like aggregates, at least in part. Using the implantation device according to this embodiment, RPE cells can be transported to a medical institution while maintaining their aggregate form and can be easily attached to a needle or cannula for use.

[0100] Cellular therapy preparations containing RPE cells are not particularly limited, but can be used, for example, to treat patients with diseases based on damage to the retinal pigment epithelium, or with atrophy or damage to the retinal pigment epithelium. Examples of diseases based on damage to the retinal pigment epithelium or with atrophy or damage to the retinal pigment epithelium include ophthalmic diseases such as age-related macular degeneration, retinitis pigmentosa and related diseases such as crystallin retinopathy, retinal pigment epithelial tears, macular dystrophy, cone-rod dystrophy, rod-cone dystrophy, macular holes, degenerative myopia, and traumatic macular disease.

[0101] As for the method of using the cell therapy preparation, for example, a cell extrusion means (e.g., a microsyringe for ophthalmic transplantation) as the first connected member is connected to the first connection part, and a cell injection means (e.g., a needle or cannula for subretinal transplantation) as the second connected member is connected to the second connection part. Then, the tip of the cell injection means is inserted into the affected area (e.g., inside the eye), and cells within the hollow fiber (e.g., RPE cells) are extruded by the cell extrusion means and transplanted to the target site (e.g., subretinal, specifically between the retina and the choroid).

[0102] Furthermore, as a method of using the cell therapy preparation, for example, a cell extrusion means (e.g., a microsyringe for ophthalmic transplantation) as the first connected member is connected to the first connection part, the tip of the injection needle (needle or cannula) of the cell culture device (transplantation device) according to this embodiment is inserted into the affected area (e.g., inside the eye), and cells (e.g., RPE cells) within the hollow fiber are extruded by the cell extrusion means and transplanted to the target site (e.g., subretinal).

[0103] This allows for easy transplantation of cells to the affected area and reduces cell loss and damage during transplantation. The above method is applicable not only to RPE cells but also to other cell types.

[0104] One embodiment of this model is a method for treating ophthalmic diseases using a cell-based pharmaceutical preparation according to this model.

[0105] One embodiment of the present invention is a method for treating an ophthalmic disease using a cell-based pharmaceutical preparation according to this embodiment, comprising the steps of: attaching a cell extrusion means (e.g., a syringe member) as a first connected member to a first connection part; attaching a cell injection means (e.g., an injection needle member) as a second connected member to a second connection part; inserting the tip of the cell injection means into the eye; and extruding and positioning cells within a hollow fiber at a target site (e.g., subretinal) using the cell extrusion means. Preferably, the step of inserting the tip of the cell injection means into the eye is the step of inserting the tip of the cell injection means attached to the cell-based pharmaceutical preparation into the eye.

[0106] One embodiment of the present invention is a method for treating an ophthalmic disease using a cell-based pharmaceutical preparation according to this embodiment, comprising the steps of: attaching a cell extrusion means (e.g., a syringe member) as a first connected member to a first connection part; inserting the tip of an injection needle (needle or cannula) of a cell culture device (implantation device) into a diseased area (e.g., inside the eye); and extruding and positioning cells within a hollow fiber at a target site (e.g., under the retina) using the cell extrusion means. Preferably, the step of inserting the tip of the cell injection means into the eye is the step of inserting the tip of the cell injection means attached to the cell-based pharmaceutical preparation into the eye.

[0107] (Example) Figure 9A is a schematic cross-sectional view illustrating a specific embodiment of the cell culture device according to this embodiment, and is a schematic cross-sectional view taken from a plane along the longitudinal direction of the hollow fiber. In Figure 9A, the cell culture device has a body member 101 that constitutes the body of the main body having an internal space 105. The cell culture device also has a first connecting member 102 at one end of the body member 101 that constitutes a first connecting part having a first connecting opening 102a, and a second connecting member 103 at the end of the body member 101 opposite to the first connecting member 102 that constitutes a second connecting part having a second connecting opening 103a. In Figure 9A, the first connecting member 102 constitutes the first connecting part and also functions as a first side wall that closes one end of the body. The second connecting member 103 constitutes the second connecting part and also functions as a second side wall that closes one end of the body. The first connecting member 102 and the second connecting member 103 are provided to close both ends of the body member 101. The first connecting portion of the first connecting member 102 is configured to be detachably attached to a first connected member (not shown) of an external element. The second connecting portion of the second connecting member 103 is configured to be detachably attached to a second connected member (not shown) of an external element. The first connecting portion of the first connecting member 102 is connectable to the first connected member so as to ensure fluid communication, and the second connecting portion of the second connecting member 103 is connectable to the second connected member so as to ensure fluid communication. The cell culture device has a single hollow fiber 104 made of a semipermeable membrane between the first connecting member 102 and the second connecting member 103 in the internal space 105 of the body member 101. The first end opening 104a of the hollow fiber 104 communicates with the first connection opening 102a of the first connecting member 102, and the second end opening 104b of the hollow fiber 104, opposite to the first end opening 104a, communicates with the second connection opening 103a of the second connecting member 13. The first connection opening 102a, the second connection opening 103a, and the cavity of the hollow fiber 104 are liquid-tightly separated from the internal space 105 of the body member 101. The body member 101 has a plurality of body openings 106 formed therein, and the internal space 105 of the body member 101 is configured to allow liquid to be supplied from the outside.

[0108] Figure 9B is a schematic side view of the cell culture device 100, observed from the side of the first connecting member 102. The first end opening 104a of the hollow fiber is located at the bottom of the first connecting opening 102a, which is located inside the first connecting portion of the first connecting member 102. A body member 101 is attached around the first connecting member 102 to ensure liquid-tightness or airtightness. Figure 9C is a schematic side view of the cell culture device, observed from the side of the second connecting member 103. The second end opening 104b of the hollow fiber is located at the bottom of the second connecting opening 103a, which is located inside the second connecting portion of the second connecting member 103. A body member 101 is attached around the second connecting member 103 to ensure liquid-tightness or airtightness.

[0109] In Figure 9A, a configuration is shown in which a plurality of body openings 106 are formed in the body member 101. However, as described above, at least one part of the body member may be made up of a mesh structure.

[0110] Furthermore, the opening of the body member 101 may be provided to be openable and closable, and for example, it may be configured so that the open or closed state can be selected by a cap or the like. The opening may also have a structure that allows it to be sealed.

[0111] In Figure 9A, the first connection opening 102a of the first connecting member 102 is divided into a proximal portion of the first connection opening on the side of the hollow fiber 104's proximal end and a distal portion of the first connection opening on the opposite side of the hollow fiber 104. The end of the hollow fiber 104 is positioned in the proximal portion of the first connection opening, and the first end-side opening 104a of the hollow fiber 104 communicates with the first connection opening 102a. Similarly, the second connection opening 103a of the second connecting member 103 is divided into a proximal portion of the second connection opening on the side of the hollow fiber 104's proximal end and a distal portion of the second connection opening on the opposite side of the hollow fiber 104. The other end of the hollow fiber 104 is positioned in the proximal portion of the second connection opening, and the second end-side opening 104b of the hollow fiber 104 communicates with the second connection opening 103a. The terms "proximal" and "distal" refer to the positional relationship when the hollow fiber 104 is grasped from its longitudinal center, with the position closer to the longitudinal center of the hollow fiber 104 being referred to as "proximal," and the position further away being referred to as "distal."

[0112] Both ends of the hollow fiber 104 are bonded to the proximal portions of the first and second connection openings, respectively, with adhesive. This bonding is carried out so that the first connection opening 102a, the second connection opening 103a, and the cavity of the hollow fiber 104 are liquid-tight (preferably airtight) separated from the internal space 105 of the body member 101.

[0113] The shapes of the first connecting portion of the first connecting member 102 and the second connecting portion of the second connecting member 103 are formed as a female Luer fitting shape and a male Luer fitting shape, respectively. Preferably, the Luer fitting shape conforms to the Luer fitting shape defined by the International Standard (ISO). Therefore, the connecting portion of the first connected member, which may be an external element, has a male Luer fitting shape corresponding to the first connecting member 102 (female Luer fitting), and the connecting portion of the second connected member, which may be an external element, has a female Luer fitting shape corresponding to the second connecting member (male Luer fitting). In one embodiment, the first connected member is a cell extrusion means (e.g., a syringe member), and the second connected member is a cell injection means (e.g., a syringe member).

[0114] In the lure fitting connection in this embodiment, a lure locking method may be employed. For example, a female threaded sleeve with threads surrounding the male lure fitting shape may be provided. For example, a configuration in which a plastic female threaded sleeve is placed around the male lure may be adopted.

[0115] The cell culture device according to this embodiment can, for example, supply culture medium to its internal space by immersing it in culture medium, and culture cells placed in the lumen of a hollow fiber.

[0116] In this specification, "cultivation" means retaining cells within the hollow fibers while supplying a culture medium to the internal space of the main body. Furthermore, in this specification, while it is desirable for cultivation to involve cell proliferation, cell proliferation is not necessarily required.

[0117] After the culture is complete and quality control is performed, the entire cell culture device is placed in a sealed container, and then a preservation solution is added to the sealed container to fill the internal space with the solution for storage and transport. Before the transplant surgery, the preservation solution of the cell culture device is replaced with a solvent suitable for use in vivo, and the cell aggregates within the hollow fibers are prepared for transplantation. For example, a cell injection means such as a subretinal injection needle is connected to the second connection point, and an extrusion syringe is connected to the first connection point. The cell aggregates within the hollow fibers can then be moved to the cell injection means by utilizing the injection pressure from liquid injection from the syringe.

[0118] Figure 10 is a schematic cross-sectional view illustrating a specific embodiment of the cell culture device according to this embodiment, and is a schematic cross-sectional view taken from a plane along the longitudinal direction of the hollow fiber. As shown in Figure 10, the cell culture device may be configured to have a liquid inlet 107 and a liquid outlet 108. This allows the culture process to be carried out while circulating the culture medium in the internal space 105. The liquid inlet 107 and liquid outlet 108 may be provided to be openable and closable, for example, by a cap or the like which the open or closed state can be selected. It is preferable that the liquid inlet 107 and liquid outlet 108 have a sealable structure. After culturing, the culture medium can be replaced with a storage solution via the liquid inlet 107 and liquid outlet 108. Then, with the liquid inlet 107 and liquid outlet 108 closed, the cell culture device can be stored and / or moved. Then, before transplantation surgery, the storage solution can be drained from the liquid outlet, and a solution that does not cause problems when it enters the body during transplantation can be injected from the liquid inlet to wash away the storage solution and perform a liquid replacement.

[0119] Figure 11 is a schematic cross-sectional view illustrating a specific embodiment of the cell culture device according to this embodiment, and is a schematic cross-sectional view taken from a plane along the longitudinal direction of the hollow fiber. As shown in Figure 11, in one embodiment, the cell culture device has a first cap 121 that is detachable from the first connection portion of the first connecting member 102. The cell culture device 100 also has a second cap 122 that is detachable from the second connection portion 103. By having the first cap 121 and the second cap 122, liquid-tight or airtight seals can be achieved between the first connection portion opening 102a, the second connection portion opening 103a, and the lumen and outside of the hollow fiber 104. As a result, when culturing cells, by immersing all or part of the cell culture device 100 in the culture medium, the culture medium can be supplied to the internal space 105 while ensuring liquid-tight or airtight seals between the first connection portion opening 102a, the second connection portion opening 103a, and the lumen and outside of the hollow fiber 104. Furthermore, the first cap 121 has a first protrusion 121a that fits into the first connection opening 102a when attached to the first connecting member 102. Also, the second cap 122 has a second protrusion 122a that fits into the second connection opening 103a when attached to the second connecting member 103. By having the first protrusion 121a that fits into the first connection opening 102a of the first cap 121, or by having the second protrusion 122a that fits into the second connection opening 103a of the second cap 122, the first end-side opening 104a or the second end-side opening 104b of the hollow fiber 104 can be sealed, effectively suppressing leakage or detachment of cells from the first end-side opening 104a or the second end-side opening 104b.

[0120] The cell culture device having the above configuration allows for the easy transfer of filamentous aggregates to other components such as injection needles. Furthermore, the cell culture device allows for cell culture within the lumen of hollow fibers composed of a semipermeable membrane, yielding filamentous aggregates. The cultured cells can be transported while contained within the hollow fibers of the cell culture device, and in some cases, the culture vessel used for cultivation can also be transported. Therefore, maintaining the quality of the cultured cells is easier, expanding the range of cell transport compared to conventional methods.

[0121] All prior art documents cited herein are incorporated herein by reference.

[0122] The upper and / or lower limits of the numerical ranges described herein can be arbitrarily combined to define a preferred range. For example, the upper and lower limits of the numerical ranges can be arbitrarily combined to define a preferred range, the upper limits of the numerical ranges can be arbitrarily combined to define a preferred range, and the lower limits of the numerical ranges can be arbitrarily combined to define a preferred range.

[0123] The claims following this disclosure are expressly incorporated herein into this disclosure, and each claim stands independently as a separate embodiment. This disclosure includes all instances in which an independent claim is replaced by its dependent claim. Furthermore, any additional embodiments derived from the independent claims and subsequent dependent claims are also expressly incorporated herein into this specification.

[0124] Although this embodiment has been described in detail above, the specific configuration is not limited to this embodiment, and any design changes that do not depart from the gist of this disclosure are also included in this disclosure.

[0125] 100: Cell culture device 101: Cell culture device 102: Cell culture device 103: Cell culture device 11: Main body 11a: First opening 11b: Second opening 12: First connection part 12a: First connection part opening 13: Second connection part 13a: Second connection part opening 14: Hollow fiber 14a: First end side opening 14b: Second end side opening 15: Internal space 16: Injection needle 17: Protective tube 17a: First protective tube 17b: Second protective tube 18: Potting resin 21: First cap 21a: First protrusion 22: Second cap 22a: Second protrusion 30: Injection needle member 31: Base 32: Injection needle 40: Syringe member 41: Tubular body 42: Tip 43: Plunger 101: Body member 102: First connecting member 102a: First connecting opening 103: Second connecting member 103a: Second connecting opening 104: Hollow fiber 104a: First end opening 104b: Second end opening 105: Internal space 106: Body opening 107: Liquid inlet 108: Liquid outlet 121: First cap 121a: First protrusion 122: Second cap 122a: Second protrusion

Claims

1. A cell culture device having a main body portion with an internal space and hollow fibers in the internal space, wherein the hollow fibers have a first end portion having a first end side opening and a second end portion located at the opposite end of the first end portion and having a second end side opening, the hollow fibers are arranged in the main body portion such that the first end side opening opens to the outside of the main body portion and the second end side opening opens to the outside of the main body portion, the lumen of the hollow fibers is liquid-tightly partitioned from the internal space, and the main body portion is configured to supply liquid from the outside to the internal space.

2. The cell culture device according to claim 1, wherein the main body portion includes a body portion, the first end side opening opens to the outside of the main body portion at one end of the body portion, and the second end side opening opens to the outside of the main body portion at the other end of the body portion.

3. The cell culture device according to claim 1, wherein the main body portion has a mesh structure at least in part.

4. The cell culture device according to claim 1, wherein the main body portion has at least one main body portion opening communicating with the internal space.

5. The cell culture device according to claim 1, wherein the main body portion has a liquid inlet and a liquid outlet respectively communicating with the internal space.

6. The cell culture device according to claim 1, wherein the hollow fiber consists of one hollow fiber.

7. The cell culture device according to claim 1, further having a first connection portion detachable from a first connected member, and the first end side opening of the hollow fiber opens to the outside at the first connection portion.

8. The cell culture device according to claim 7, wherein the first connection portion is connectable to the first connected member such that the inside of the first connected member and the first end side opening are spatially connected.

9. The cell culture device according to claim 7, wherein the first connection portion has a female Luer fitting shape.

10. The cell culture device according to claim 9, wherein the connection portion of the first connected member has a male Luer fitting shape corresponding to the female Luer fitting shape of the first connection portion.

11. The cell culture device according to claim 7, wherein the first connected member is a cell extrusion means (for example, a syringe member).

12. The cell culture device according to claim 7, further comprising a second connecting portion detachably attached to a second connecting member at a portion different from the first connecting portion, wherein the second end-side opening of the hollow fiber opens to the outside at the second connecting portion.

13. The cell culture device according to claim 12, wherein the second connecting portion is connectable to the second connecting member such that the inside of the second connecting member and the second end-side opening are spatially in communication.

14. The cell culture device according to claim 12, wherein the second connecting portion has a male Luer fitting shape.

15. The cell culture device according to claim 14, wherein the connecting portion of the second connecting member has a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion.

16. The cell culture device according to claim 12, wherein the second connecting member is a cell injection means (for example, an injection needle member).

17. The cell culture device according to claim 12, further comprising a protective tube made of metal or resin surrounding the outer periphery of the second end of the hollow fiber.

18. The cell culture device according to claim 17, wherein the protective tube protrudes to the outside from the second connecting portion.

19. The cell culture device according to claim 18, wherein the second connecting portion has a male Luer fitting shape, the second connecting member is an injection needle member including a base portion having a female Luer fitting shape corresponding to the male Luer fitting shape of the second connecting portion and an injection needle disposed at the tip of the base portion, and the protective tube protrudes to the outside from the second connecting portion such that the tip portion of the protective tube contacts the base-side end portion of the injection needle when the second connecting portion and the second connecting member are connected.

20. The cell culture device according to claim 7, further comprising an injection needle at a portion different from the first connecting portion of the main body portion, wherein the injection needle is installed such that fluid communication between the lumen of the injection needle and the second end-side opening is ensured.

21. The cell culture device according to claim 20, wherein the injection needle is installed such that at least a part of the second end of the hollow fiber enters the lumen of the injection needle.

22. The cell culture device according to claim 20, wherein the injection needle has a tapered shape at least in part, whereby the tip of the injection needle is thinner.

23. A cell therapy preparation comprising a cell culture device and a transplant material containing cells according to any one of claims 1 to 22, wherein the transplant material is disposed within the lumen of the hollow fiber.

24. The cell therapy preparation according to claim 23, wherein the cells are at least one selected from stem cells, progenitor cells, somatic cells, and cells differentiated from stem cells or progenitor cells.

25. The cell therapy preparation according to claim 23, wherein the cells include retinal pigment epithelial cells.

26. The cell therapy preparation according to claim 23, wherein at least a portion of the cells form aggregates within the lumen of the hollow fiber.

27. A method for producing a cell-based pharmaceutical preparation according to claim 23, comprising the steps of: placing the cells in the lumen of the hollow fiber; and placing a culture medium in the internal space of the main body and culturing the cells.

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