WO2023127132A1 - Container for cell culture use - Google Patents

Container for cell culture use Download PDF

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Publication number
WO2023127132A1
WO2023127132A1 PCT/JP2021/048882 JP2021048882W WO2023127132A1 WO 2023127132 A1 WO2023127132 A1 WO 2023127132A1 JP 2021048882 W JP2021048882 W JP 2021048882W WO 2023127132 A1 WO2023127132 A1 WO 2023127132A1
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WO
WIPO (PCT)
Prior art keywords
film
cell culture
space
plate body
internal space
Prior art date
Application number
PCT/JP2021/048882
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French (fr)
Japanese (ja)
Inventor
学司 加藤
順一 桑原
Original Assignee
株式会社サンプラテック
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社サンプラテック filed Critical 株式会社サンプラテック
Priority to PCT/JP2021/048882 priority Critical patent/WO2023127132A1/en
Priority to JP2023570608A priority patent/JPWO2023127132A1/ja
Publication of WO2023127132A1 publication Critical patent/WO2023127132A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present disclosure relates to a cell culture vessel for culturing cells, and in particular, a cell culture vessel used for perfusion culture that automatically supplies culture medium at a constant low speed and discharges the same amount of culture medium at the same time. Regarding.
  • the cells are transferred to a new container according to the growth of the cells. At that time, contamination is likely to occur in the process of transferring to a larger container or increasing the number of containers. Therefore, cell culture using conventional culture vessels such as dishes, well plates, and culture flasks has a high risk of contamination.
  • the present disclosure has been conceived under such circumstances, and it is possible to continue cell perfusion culture by shifting to expansion culture with an expanded volume without transferring the cells to another container.
  • the main object is to provide a container for cell culture that is capable of
  • a cell culture vessel includes a first bag portion including a first film-like member positioned on one side in a first direction, and a first plate body attached to the first film-like member; a second film-like member located on the other side in the first direction with respect to the first bag portion and having an outer peripheral portion connected to the outer peripheral portion of the first film-like member; a suction member attached to one plate body, the suction member facing the other side in the first direction and having an annular suction pad capable of being in close contact with the second bag portion; One side end of each bag portion communicates with the outside, and the other side end is inside the adsorption member when viewed in the first direction, and is connected to the first bag portion and the second bag portion in the first direction.
  • first flow path and a second flow path located in the central internal space between the central internal space and the first direction when the suction pad is in close contact with the second bag portion.
  • the central interior space and the peripheral interior space are in communication.
  • the first film-like member has an annular shape with a first opening on the inner peripheral side, and the first plate body closes the first opening.
  • the second bag portion includes a second plate body separate from the second film-like member, the second plate body faces one side in the first direction, and It has a substantially flat first bottom surface, and the suction pad can be in close contact with the first bottom surface.
  • the second bag part includes a second plate body attached to the second film-like member, the second plate body faces one side in the first direction, and substantially It has a flat first bottom surface, and the suction pad can be brought into close contact with the first bottom surface.
  • the second film-like member has an annular shape with a second opening on the inner peripheral side, and the second plate body closes the second opening.
  • said second bag part comprises a second plate body attached to said second film-like member, said second plate body being attached to said second film-like member by said first film-like member. located on the other side of the direction and overlapping the central interior space when viewed in the first direction.
  • the area of the second plate body is larger than that of the first plate body when viewed in the first direction.
  • the adsorption member has at least one suction path that penetrates the first plate body in the first direction, one side end of the suction path communicates with the outside, and The other end communicates with the inside of the suction pad.
  • the first film-like member is provided with a plurality of ports each of which has one side end communicating with the outside and the other side end communicating with the surrounding internal space.
  • the first plate body includes a first annular plate portion having an annular shape when viewed in the first direction, and closing an inner side of the first annular plate portion when viewed in the first direction, and a bulging portion projecting to one side in the first direction, a gas permeable membrane is attached to the first annular plate portion, and the central internal space is formed between the second bag portion and the
  • the first plate body includes a first space sandwiched between the gas permeable membrane and the first annular plate portion, and a second space surrounded by the gas permeable membrane and the bulging portion. It has a plurality of third flow paths with one side end communicating with the outside and the other side end communicating with the second space.
  • the first plate body is provided with a liquid permeable membrane filter, and the central inner space is located on the other side in the first direction with respect to the membrane filter. and a third space located on one side in the first direction, the other side end of the first flow path communicates with the first space, and the other side of the second flow path The end communicates with the third space.
  • a portion of the first plate body that forms the third space is formed with a ceiling recess that is recessed toward one side in the first direction, and the ceiling recess is formed in the second space. connected to the stream.
  • a plurality of densely arranged suction pads are provided in a region of the surface facing one side in the first direction that includes the inner side of the suction pad when viewed in the first direction. is provided.
  • a third film-like member having an outer peripheral portion connected to the outer peripheral portion of the second film-like member is further provided, and the other side of the second bag portion in the first direction is connected to the third film-like member. and a communication means for communicating between the central internal space and the fourth space when the suction pad is not in close contact with the second bag portion.
  • FIG. 1 is a perspective view showing a cell culture container according to a first embodiment of the present disclosure
  • FIG. FIG. 2 is a plan view of the cell culture vessel shown in FIG. 1
  • 3 is a cross-sectional view taken along line III-III of FIG. 2
  • FIG. 3 is a cross-sectional view taken along line IV-IV of FIG. 2
  • FIG. FIG. 3 is a cross-sectional view taken along line VV of FIG. 2
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure
  • Fig. 10 is a perspective view showing a cell culture container according to a second embodiment of the present disclosure
  • FIG. 11 is a plan view of the culture vessel shown in FIG. 10
  • 12 is a cross-sectional view along line XII-XII of FIG. 11
  • FIG. FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a third embodiment of the present disclosure
  • FIG. 10 is a diagram showing an arrangement state of a plurality of recesses formed on the first bottom surface of the second plate body, viewed in the first direction;
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a fourth embodiment of the present disclosure;
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a fifth embodiment of the present disclosure;
  • FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a sixth embodiment of the present disclosure;
  • FIG. 4 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a seventh embodiment of the present disclosure;
  • FIG. 4 is a cross-sectional view, similar to FIG.
  • FIG. 4 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a ninth embodiment of the present disclosure
  • FIG. 10 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a tenth embodiment of the present disclosure
  • FIG. 11 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to an eleventh embodiment of the present disclosure
  • FIG. 12 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a twelfth embodiment of the present disclosure
  • FIG. 4 is a plan view showing a modification of the cell culture vessel according to the present disclosure
  • FIG. 25 is a plan view showing a usage example of the cell culture vessel shown in FIG. 24
  • FIG. 25 is a plan view showing a usage example of the cell culture vessel shown in FIG. 24;
  • FIG. 1 is a perspective view of a cell culture vessel A1.
  • FIG. 2 is a plan view of the cell culture vessel A1.
  • the first film-like member 1 and the second film-like member 2 each form an annular shape (in this embodiment, an annular shape) and overlap each other when viewed in the first direction x. .
  • the first film-like member 1 and the second film-like member 2 are in the form of thin films having flexibility, and are easily flexurally deformed following an external force.
  • the first film-like member 1 is positioned on one side in the first direction x (hereinafter referred to as “first direction one side x1”), and the second film-like member 2 is positioned on the other side in the first direction x ( hereinafter referred to as “first direction other side x2”).
  • the first film-like member 1 has a first opening 11 on the inner peripheral side
  • the second film-like member 2 has a second opening 21 on the inner peripheral side.
  • the outer peripheral portion 12 of the first film-like member 1 and the outer peripheral portion 22 of the second film-like member 2 are joined by an appropriate means such as thermal welding and connected to each other.
  • the constituent material of the first film-like member 1 and the second film-like member 2 is not particularly limited, and examples thereof include polyethylene, polyolefin, styrene-based elastomer, and thermoplastic elastomer.
  • the first film-like member 1 and the second film-like member 2 are not limited to the structure in which the outer peripheral portions 12 and 22 are joined by heat welding as described above. It may be a molded product formed in the
  • a plurality of ports 19 are provided at appropriate locations on the first film member 1 . As shown in FIGS. 3 and 4 , port holes 13 are formed in the first film-like member 1 at locations corresponding to the respective ports 19 . The multiple ports 19 will be described later.
  • the first plate body 3 is located on the first direction one side x1 with respect to the second plate body 4. As shown in FIGS. The first plate body 3 closes the first opening 11 of the first film member 1 .
  • the second plate body 4 is positioned on the other side x2 in the first direction with respect to the first plate body 3 .
  • the second plate body 4 closes the second opening 21 of the second film member 2 .
  • the first plate body 3 includes a first annular plate portion 31, a second annular plate portion 32, a bulging portion 33 and a filter retainer .
  • the first annular plate portion 31 has an annular shape when viewed in the first direction x, and includes a substantially annular plate portion in the present embodiment.
  • the first annular plate portion 31 has a first flow path 311 and an adsorption member groove 312 .
  • the adsorption member concave groove 312 is an annular groove surrounded by an annular tubular portion extending in the first direction other side x2, and is a portion for disposing the adsorption member 5 .
  • the first channel 311 will be described later.
  • the bulging portion 33 is connected to the inner peripheral edge of the first annular plate portion 31 .
  • the bulging portion 33 closes the inside of the first annular plate portion 31 when viewed in the first direction x.
  • the bulging portion 33 protrudes toward the first direction one side x1 with respect to the first annular plate portion 31 .
  • the bulging portion 33 has a plurality of third flow paths 331 .
  • the 3rd flow path 331 is mentioned later.
  • the second annular plate portion 32 is connected to the outer peripheral edge of the first annular plate portion 31 via a stepped portion, and is an annular plate-like portion.
  • the inner peripheral portion of the first film member 1 is joined to the lower surface of the outer peripheral portion of the second annular plate portion 32 (the surface facing the other side x2 in the first direction) by, for example, ultrasonic welding or adhesion. ing. Thereby, the first plate body 3 closes the first opening 11 of the second film member 2 .
  • the first annular plate portion 31, the second annular plate portion 32, and the bulging portion 33 are made of a hard material, such as a resin molded product.
  • the constituent materials of the first annular plate portion 31, the second annular plate portion 32, and the bulging portion 33 are not particularly limited, and examples thereof include polystyrene, polycarbonate, and COP (cycloolefin polymer).
  • the filter presser 34 is a member that sandwiches the membrane filter 7 and is fitted into the cylindrical portion of the first annular plate portion 31 .
  • the filter retainer 34 has a cylindrical portion 341 , a recessed ceiling portion 342 and a second flow path 343 .
  • the cylindrical portion 341 has a cylindrical shape and is fitted into the cylindrical portion of the first annular plate portion 31 .
  • the ceiling concave portion 342 is connected to the tubular portion 341 and closes the inside of the tubular portion 341 .
  • the ceiling recessed portion 342 is recessed in the first direction one side x1.
  • the first annular plate portion 31 and the filter retainer 34 (cylindrical portion 341) sandwich the membrane filter 7 from both sides in the first direction x.
  • the filter retainer 34 is made of a hard material, such as a resin molded product.
  • the constituent material of the filter retainer 34 is not particularly limited, and examples thereof include polystyrene, polycarbonate, COP, and the like.
  • the first film-like member 1 and the first plate body 3 described above respectively correspond to constituent elements of the "first bag portion".
  • the constituent elements of the first bag portion (the first film-like member 1 and the first plate body 3) are appropriately denoted by B1.
  • the second plate body 4 has a substantially disc shape and includes a main plate portion 41 .
  • the main plate portion 41 is flat and has a first bottom surface 41a.
  • the first bottom surface 41a faces the first direction one side x1 and is substantially flat.
  • the first bottom surface 41a being "substantially flat” means that the flatness is enough to allow the suction pads 52 of the suction member 5 described later to be in close contact, and does not include a shape such as a curved shape that makes it difficult for the suction pads 52 to be in close contact. do.
  • the first bottom surface 41a is a smooth plane.
  • the inner portion of the second film-like member 2 is applied, for example, by a technique such as heat sealing or adhesion.
  • the perimeter is joined.
  • the second plate body 4 closes the second opening 21 of the second film member 2 .
  • the second plate body 4 is made of a hard material, such as a resin molded product.
  • the constituent material of the second plate body 4 is not particularly limited, and examples thereof include polystyrene, polycarbonate, and COP.
  • the inner peripheral portion of the second film member 2 is joined to the upper surface of the outer peripheral portion of the second plate member 4 (the first bottom surface 41a facing the one side x1 in the first direction).
  • the inner peripheral portion of the second film member 2 may be joined to the lower surface of the outer peripheral portion of the two-plate body 4 (the surface facing the other side x2 in the first direction).
  • the second film-like member 2 and the second plate body 4 described above each correspond to constituent elements of the "second bag portion".
  • the constituent elements of the second bag portion (the second film-like member 2 and the second plate body 4) are appropriately denoted by B2.
  • the adsorption member 5 shown in FIGS. 3 to 5 is for isolating the internal space of the cell culture container A1 into the central internal space S1 and the peripheral internal space S2.
  • the adsorption member 5 has an annular shape as a whole when viewed in the first direction x.
  • the adsorption member 5 is attached to the first plate body 3 .
  • the adsorption member 5 has a block portion 51 , an adsorption pad 52 and a suction port 53 .
  • a large portion of the block portion 51 is accommodated in the suction member recessed groove 312 .
  • the block portion 51 is press-fitted into the suction member recessed groove 312 and fixed by adhesion or the like as necessary.
  • the suction pad 52 is an annular suction cup connected to the first direction other side x2 of the block portion 51 and facing the first direction other side x2. More specifically, the suction pad 52 has a pair of annular lip portions extending both radially outward and radially inward from the lower end of the block portion 51 (the other side x2 end in the first direction). The suction pad 52 can be brought into close contact with the first bottom surface 41a of the second plate body 4, as shown in FIG.
  • the central internal space S1 and the surrounding internal space S2 are cut off.
  • the central internal space S1 is the inside of the adsorption member 5 when viewed in the first direction x, and the first film-like member 1 (first bag portion B1) and the second film-like member 2 (second bag portion) in the first direction x. B2).
  • the surrounding internal space S ⁇ b>2 is located outside the adsorption member 5 when viewed in the first direction x and is surrounded by the first film member 1 , the second film member 2 and the adsorption member 5 .
  • the suction port 53 is for connecting suction means (not shown) such as a pump to suck the inside of the suction pad 52 .
  • the suction port 53 has an extension 531 , a suction path 532 and a protrusion 533 .
  • the extending portion 531 has a cylindrical shape extending from the block portion 51 in the first direction one side x1 and has an outer peripheral surface with a circular cross section. The extending portion 531 protrudes from the first annular plate portion 31 to the first direction one side x1 through the through hole 313 formed in the first annular plate portion 31 .
  • a projecting portion 533 that protrudes outward from the outer peripheral surface of the extending portion 531 is provided at the tip of the extending portion 531 (first direction one side x1 end).
  • the inner diameter dimension of the through hole 313 is slightly smaller than the outer diameter dimension of the extending portion 531 in the natural state.
  • the extending portion 531 is press-fitted into the through hole 313 .
  • the through hole 313 (the first annular plate portion 31) is in close contact with (contacts with) the outer peripheral surface of the extending portion 531 to surround the outer peripheral surface.
  • the protrusion 533 can come into contact with the upper end (first direction one side x1 end) of the first annular plate portion 31 .
  • the suction path 532 penetrates the first plate body 3 in the first direction x.
  • One side end (first direction one side x1 end) of the suction path 532 communicates with the outside, and the other side end (first direction other side x2 end) communicates with the inside of the suction pad 52 .
  • the adsorption member 5 is made of, for example, a soft material that is easily elastically deformable.
  • the constituent material of the adsorption member 5 is not particularly limited, and examples thereof include silicone rubber and thermoplastic elastomer.
  • the gas permeable membrane 6 shown in FIGS. 3 to 5 is a thin membrane having gas permeability.
  • the gas-permeable membrane 6 is impermeable to liquid and impermeable to liquid.
  • the gas permeable membrane 6 is attached to the first annular plate portion 31 of the first plate body 3 .
  • the gas permeable film 6 is fixed in close contact with the lower surface of the first annular plate portion 31 (the surface facing the other side x2 in the first direction) by an appropriate means such as adhesion.
  • the central internal space S1 includes a first space S11 and a second space S12 that are separated from each other.
  • the first space S ⁇ b>11 is sandwiched between the second plate body 4 (first bottom surface 41 a ), the gas permeable film 6 and the first annular plate portion 31 .
  • the second space S ⁇ b>12 is surrounded by the gas permeable membrane 6 and the bulging portion 33 .
  • the constituent material of the gas permeable membrane 6 is not particularly limited, and examples thereof include a hydrophobic filter made of PET (polyester) or PC (polycarbonate).
  • the membrane filter 7 shown in FIG. 3 is a liquid-permeable membrane through which liquid can pass.
  • the membrane filter 7 can pass a liquid such as a culture solution. Gases such as air can also pass through the membrane filter 7 .
  • non-liquid contents such as cultured cells cannot pass through the membrane filter 7 and are captured.
  • the central internal space S1 includes a partitioned third space S13.
  • the third space S13 is positioned on the one side x1 in the first direction with respect to the membrane filter 7 .
  • the third space S13 is surrounded by the membrane filter 7 and the filter retainer .
  • the first space S11 is positioned on the other side x2 in the first direction with respect to the membrane filter 7 .
  • the constituent material of the membrane filter 7 is not particularly limited, and examples thereof include hydrophilic filters made of PET (polyester) or PC (polycarbonate).
  • the first flow path 311, the second flow path 343 and the plurality of third flow paths 331 are cylindrical communicating paths that communicate with the outside and the internal space of the cell culture vessel A1.
  • Each of the first flow path 311 and the second flow path 343 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) to the central internal space S1.
  • the other side end (first direction other side x2 end) of the first flow path 311 communicates with the first space S11.
  • the other side end (first direction other side x2 end) of the second flow path 343 communicates with the third space S13.
  • a ceiling recessed portion 342 of the filter retainer 34 that defines the third space S13 is connected to a second flow path 343 .
  • Each of the plurality of third flow paths 331 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) communicated with the second space S12.
  • Each of the plurality of ports 19 shown in FIGS. 3 and 4 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) communicates with the outside. 1 It communicates with the surrounding internal space S2 through the port hole 13 of the film-like member 1 .
  • the cell culture vessel A1 is used for perfusion culture in which the culture medium is automatically supplied at a constant low speed at all times, and the same amount of culture medium is discharged outside the vessel at the same time.
  • the cultured cells and the culture solution to be accommodated in the cell culture vessel A1 are not particularly limited.
  • the adsorption pad 52 of the adsorption member 5 is brought into close contact with the first bottom surface 41a of the second plate body 4 in advance, as shown in FIG.
  • the first bottom surface 41a is smooth and can be suitably used as a culture surface for adherent cells.
  • the suction pad 52 of the suction member 5 attached to the first plate body 3 can be brought into close contact with the first bottom surface 41a.
  • the central internal space S1 and the surrounding internal space S2 are cut off.
  • the inside of the suction pad 52 is sucked by suction means (not shown) connected to the suction port 53 (suction path 532) shown in FIG. power is increased. As a result, the central internal space S1 and the surrounding internal space S2 are more reliably cut off.
  • the culture solution is supplied to the cell culture vessel A1 and the air is discharged through the first channel 311 and the second channel 343 to fill the central internal space S1 with the culture solution.
  • a tube 81 is connected to each of the first flow path 311 and the second flow path 343 , and one end of the first flow path 311 (first direction one side x1 end) is connected via the tube 81 .
  • a perfusion pump (not shown) is connected to the second channel 343
  • a waste liquid container (not shown) is connected to one end of the second flow path 343 (first direction one side x1 end) via a tube 81 .
  • the cultured cells are supplied to the central internal space S1 together with the culture solution.
  • the other side end (first direction other side x2 end) of the first flow path 311 communicates with the first space S11 of the central internal space S1.
  • the first space S11 is filled with the culture medium supplied through the first channel 311 .
  • Cultured cells supplied via the first channel 311 also flow into the first space S11.
  • the air in the first space S11 passes through the gas-permeable membrane filter 7, passes through the third space S13, and is discharged from the second flow path 343 to the outside.
  • the arrangement of the first flow path 311 and the second flow path 343 provided in the first plate body 3 is not particularly limited.
  • the first flow path 311 and the second flow path 343 are spaced apart from each other on opposite sides of the central portion of the first plate body 3 when viewed in the first direction x. According to such a configuration, the culture solution can be uniformly circulated to the central internal space S1 (first space S11).
  • Liquid can pass through the membrane filter 7 located on the first direction one side x1 of the first space S11. Therefore, the culture solution filling the first space S11 passes through the membrane filter 7 and flows into the third space S13. 7 shows a state in which the first space S11 and the third space S13 are filled with the culture solution C.
  • the filter presser 34 forming the third space S13 located above the membrane filter 7 (first direction one side x1) is formed with a ceiling recess 342 recessed in the first direction one side x1. It is connected to the second channel 343 .
  • air bubbles in the first space S11 pass through the membrane filter 7 and enter the third space S13. Moving. By operating the perfusion pump in this state, air bubbles remaining in the third space S13 can move into the second channel 343 through the ceiling recess 342 and be discharged to the outside of the cell culture vessel A1.
  • a new culture solution C is supplied to the first space S11 (central internal space S1) through the first channel 311, and is supplied to the third space S13 (central internal space S1) through the second channel 343.
  • the culture solution C in S1) is discharged to the outside.
  • the culture medium C in the cell culture vessel A1 (the central internal space S1) is gradually replaced.
  • the cultured cells float against their own weight and are almost never attracted to the membrane filter 7 provided on the side of the recess 342 on the ceiling. Cultured cells in the culture solution C cannot pass through the membrane filter 7 even if they reach the membrane filter 7 . As a result, even if the cultured cells float in the culture solution C in the first space S11, the cultured cells in the culture solution C are prevented from being unduly discharged from the second channel 343 .
  • a gas permeable membrane 6 is attached to the first annular plate portion 31 of the first plate body 3 .
  • the central internal space S1 is surrounded by a first space S11 sandwiched between the first bottom surface 41a of the second plate body 4, the gas permeable membrane 6 and the first annular plate portion 31, the gas permeable membrane 6 and the bulging portion 33. and a second space S12.
  • the bulging portion 33 (first plate body 3) has a plurality of third flow paths 331, and one side end (first direction one side x1 end) of each third flow path 331 communicates with the outside.
  • the side end (first direction other side x2 end) communicates with the second space S12.
  • the second space S12 is maintained in a predetermined gas atmosphere by supplying a predetermined gas to the bulging portion 33 through the plurality of third flow paths 331 and discharging the predetermined gas as necessary. can do.
  • the contents of the first space S11 (the culture medium C and the cultured cells) are maintained in a ventilated state with the second space S12 through the gas permeable membrane 6, and the gas in the second space S12 can be taken in. is.
  • the type of gas supplied to the third flow path 331 the gas atmosphere of the content gas in the first space S11 can be changed easily and quickly in the second space S12. For example, compared with the case of creating a predetermined gas atmosphere using an incubator or the like, it is possible to ventilate a desired gas atmosphere with a simple configuration.
  • a cell culture vessel A1 of the present embodiment includes a first film-like member 1 and a second film-like member 2 .
  • the first film-like member 1 and the second film-like member 2 are each annular, and the outer peripheral portions 12 and 22 are connected to each other. 2 It is blocked by the plate body 4 .
  • the suction pad 52 of the suction member 5 When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , and the central internal space S1 (first space S11) filled with the culture medium C is cut off.
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the storage space for the culture solution C (the central internal space S1
  • the volumes of (the first space S11) and the surrounding internal space S2) are expanded to a large capacity (see FIG. 9).
  • the cell culture container A1 of the present embodiment it is possible to shift to expansion culture in the middle of cell culture (perfusion culture) and continue cell culture (perfusion culture) without moving the container. be. Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the first film-like member 1 is provided with a plurality of ports 19 .
  • One side end (first direction one side x1 end) of each port 19 communicates with the outside, and the other side end (first direction other side x2 end) communicates with the surrounding internal space S2.
  • a tube 82 is connected to each of the plurality of ports 19, and a solution injection pump (not shown) is connected to some of the ports 19 via the tube 82.
  • the remaining port 19 is connected via a tube 82 to a waste liquid discharge pump and further via a tube 82 to a waste liquid container (not shown).
  • one of the plurality of ports 19 used for discharge is connected to a waste liquid discharge port 84 via a tube 83 toward the surrounding internal space S2.
  • the waste liquid outlet 84 is provided with a filter 85 (the tube 83, the waste liquid outlet 84 and the filter 85 are omitted in the drawings other than FIG. 9).
  • the supply amount and discharge amount of the culture solution C are appropriately adjusted.
  • the central internal space S1 and the peripheral internal space S2 are not filled with the culture solution C, and the position of the liquid surface of the culture solution C in the cell culture vessel A1 is adjusted. Therefore, in the first direction one side x1 of the central internal space S1 and the peripheral internal space S2, there is a space where the culture solution C does not exist.
  • the third channel 331 was used for the gas atmosphere adjustment at the stage of performing perfusion culture only in the central internal space S1 (first space S11), the gas atmosphere adjustment at the stage in which the peripheral internal space S2 was communicated was This can be done using the first channel 311 and the second channel 343 .
  • the culture solution C can be injected into and discharged from the surrounding internal space S2 through the plurality of ports 19 as described above, the adhesion of the suction pad 52 to the first bottom surface 41a as shown in FIG.
  • An appropriate amount of culture medium C can be supplied to the surrounding internal space S2 through the port 19 in advance while maintaining the state.
  • the suction pad 52 is released from the adhered state after that, the culture medium C is prevented from suddenly flowing into the surrounding internal space S2, and turbulence caused by the rapid flow of the culture medium C can be suppressed. . This contributes to proper transfer to expansion culture.
  • the suction state of the suction pad 52 can be quickly and easily removed. can be released. This makes it possible to automate the expansion of the volume of the culture vessel with an extremely simple mechanism.
  • ⁇ Second embodiment> 10 to 12 show a cell culture vessel according to a second embodiment of the present disclosure.
  • the same or similar elements as those of the cell culture vessel A1 of the above embodiment are denoted by the same reference numerals as those of the above embodiment, and description thereof will be omitted as appropriate.
  • the cell culture vessel A2 of this embodiment differs from the cell culture vessel A1 of the above embodiment mainly in the structure of the adsorption member 5.
  • the adsorption member 5 further has a columnar portion 54 .
  • the columnar portion 54 is arranged away from the suction port 53 , and in the illustrated example, is provided at the farthest position from the suction port 53 in the circumferential direction of the adsorption member 5 .
  • the columnar portion 54 has a columnar shape extending from the block portion 51 in the first direction one side x1 and has an outer peripheral surface with a circular cross section.
  • the columnar portion 54 protrudes from the first annular plate portion 31 to the first direction one side x1 through a through hole 314 formed in the first annular plate portion 31 .
  • a protrusion 541 that protrudes outward from the outer peripheral surface of the columnar portion 54 is provided at the tip of the columnar portion 54 (first direction one side x1 end). .
  • the inner diameter of the through hole 314 is slightly smaller than the outer diameter of the columnar portion 54 in the natural state.
  • the columnar portion 54 is press-fitted into the through hole 314 .
  • the through hole 314 (the first annular plate portion 31) is in close contact with (contacts with) the outer peripheral surface of the columnar portion 54 to surround the outer peripheral surface.
  • the projecting portion 541 overlaps with the projecting portion 541 .
  • the projecting portion 541 is adjacent above the first annular plate portion 31 . Thereby, the protrusion 541 can come into contact with the upper end (first direction one side x1 end) of the first annular plate portion 31 .
  • the first film-like member 1 and the second film-like member 2 are each annular, the outer peripheral portions 12 and 22 are connected to each other, and the first opening 11 on the inner peripheral side and the second film-like member 2 are connected to each other.
  • Two openings 21 are blocked by the first plate body 3 and the second plate body 4 .
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the adsorption member 5 has a columnar portion 54, and at the tip of the columnar portion 54 (first direction one side x1 end), there is a protrusion projecting outward from the outer peripheral surface of the columnar portion 54. 541 is provided.
  • the projecting portion 541 is adjacent to the upper side of the first annular plate portion 31 and can abut on the upper end of the first annular plate portion 31 .
  • the adsorption member 5 will not move to the first bottom surface 41a (the first bottom surface 41a) of the first plate body 3 A pulling force acts downward (the second side in the first direction x2) on the 2 plate body 4) side. Even if a downward force acts on the attracting member 5 in this manner, the projecting portion 541 is locked by the upper end of the first annular plate portion 31 , so that the attracting member 5 is separated from the first plate body 3 . is prevented.
  • a projection portion 533 projecting outward from the outer peripheral surface of the extension portion 531 is provided at the tip of the extension portion 531 (first direction one side x1 end). It is As a result, even if a downward force acts on the attracting member 5 , the extending portion 531 is locked by the upper end of the first annular plate portion 31 , so that the attracting member 5 is separated from the first plate body 3 . is prevented.
  • the adsorption member 5 has one columnar portion 54 in this embodiment, it may have a plurality of columnar portions 54 . When the adsorption member 5 has a plurality of columnar portions 54 , it is preferable that the extension portion 531 and the plurality of columnar portions 54 are evenly distributed in the circumferential direction of the adsorption member 5 .
  • the cell culture vessel A2 of the present embodiment has the same effects as the cell culture vessel A1 of the above embodiment.
  • ⁇ Third Embodiment> 13 and 14 show a cell culture vessel according to a third embodiment of the present disclosure.
  • the cell culture vessel A3 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second plate body 4 .
  • a plurality of recesses 411 are formed in the first bottom surface 41a of the second plate body 4 (main plate portion 41).
  • the plurality of recesses 411 are formed on the first bottom surface 41a so as to include regions inside the suction pads 52 when viewed in the first direction x.
  • the plurality of recesses 411 are densely arranged along the in-plane direction of the first bottom surface 41a.
  • the plurality of recesses 411 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
  • the first film-like member 1 and the second film-like member 2 are each annular, the outer peripheral portions 12 and 22 are connected to each other, and the first opening 11 on the inner peripheral side and the second film-like member 2 are connected to each other.
  • Two openings 21 are blocked by the first plate body 3 and the second plate body 4 .
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • a plurality of recesses 411 are provided on the first bottom surface 41a of the main plate portion 41 (second plate body 4).
  • a container bottom (first bottom surface 41a) including a plurality of recesses 411 may be coated with a non-adherent cell coating that favors spheroid formation.
  • cells are three-dimensionally aggregated, and spheroids are likely to be formed.
  • air bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the concave portion 411, and the membrane filter 7 provided on the ceiling concave portion 342 side prevents the spheroids from is not discharged.
  • the cells (spheroids) perfused and cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture with the peripheral internal space S2 communicated without being removed from the cell culture container A3. becomes possible.
  • the cell culture vessel A3 of the present embodiment has the same effects as the cell culture vessel A1 of the above embodiment.
  • FIG. 15 shows a cell culture container according to a fourth embodiment of the present disclosure.
  • the cell culture vessel A4 of the present embodiment differs from the cell culture vessel A1 of the above embodiment mainly in the configuration of the second bag portion B2.
  • the second film-like member 2 does not have the second opening 21 and has a substantially circular shape.
  • the second plate body 4 has a substantially annular shape.
  • the second plate body 4 includes an annular main plate portion 41 .
  • the main plate portion 41 has a first bottom surface 41a.
  • the first bottom surface 41a faces the first direction one side x1 and is substantially flat.
  • the first bottom surface 41a is a portion to which the suction pad 52 is brought into close contact.
  • the lower surface (the surface facing the other side x2 in the first direction) of the second plate body 4 (the main plate portion 41) is joined to the surface facing the one side x1 in the first direction in the central portion of the second film member 2.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the cell culture vessel A4 of the present embodiment has the same effects as those of the above embodiment within the same configuration range as the cell culture vessel A1 of the above embodiment.
  • FIG. 16 shows a cell culture container according to a fifth embodiment of the present disclosure.
  • the cell culture vessel A5 of this embodiment differs from the cell culture vessel A4 of the above embodiment in the configuration of the second bag portion B2.
  • the second plate body 4 is not provided.
  • the suction pad 52 can be brought into close contact with the bottom surface 23a of the second film member 2 facing the one side x1 in the first direction.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11).
  • the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate.
  • the cell culture vessel A4 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the cell culture vessel A5 of the present embodiment has the same effects as those of the above embodiment within the same range of configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 17 shows a cell culture container according to a sixth embodiment of the present disclosure.
  • the cell culture vessel A6 of the present embodiment differs from the cell culture vessel A4 of the above embodiment in the configuration of the second film-like member 2 .
  • the configuration of the second film member 2 is different from that of the cell culture vessel A4.
  • a plurality of recesses 231 are formed in the bottom surface 23a of the second film member 2 facing the first direction one side x1.
  • the plurality of recesses 231 are formed including areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 23a.
  • the plurality of recesses 231 are densely arranged along the in-plane direction of the bottom surface 23a in the same manner as the plurality of recesses 411 shown in FIG.
  • the plurality of recesses 231 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) is shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the bottom surface 23a of the second film member 2 is provided with a plurality of recesses 231.
  • a container bottom (bottom surface 23a) including a plurality of recesses 231 may be coated with a non-adhesive cell coating that favors spheroid formation.
  • cells are three-dimensionally aggregated, and spheroids are likely to be formed.
  • bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the recess 231, and the membrane filter 7 provided on the ceiling recess 342 side prevents spheroids from is not discharged.
  • the cells (spheroids) that have been perfusion cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture in which the surrounding internal space S2 is communicated without being removed from the cell culture container A6. becomes possible.
  • the cell culture vessel A6 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 18 shows a cell culture container according to a seventh embodiment of the present disclosure.
  • the cell culture vessel A7 of this embodiment differs from the cell culture vessel A5 of the above embodiment in the configuration of the second film member 2 .
  • a plurality of recesses 231 are formed on the bottom surface 23a of the second film-like member 2 facing the one side x1 in the first direction.
  • the plurality of recesses 231 are formed including areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 23a.
  • the plurality of recesses 231 are densely arranged along the in-plane direction of the bottom surface 23a in the same manner as the plurality of recesses 411 shown in FIG.
  • the plurality of recesses 231 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11).
  • the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate.
  • the cell culture vessel A7 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the bottom surface 23a of the second film member 2 is provided with a plurality of recesses 231.
  • a container bottom (bottom surface 23a) including a plurality of recesses 231 may be coated with a non-adhesive cell coating that favors spheroid formation.
  • cells are three-dimensionally aggregated, and spheroids are likely to be formed.
  • bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the recess 231, and the membrane filter 7 provided on the ceiling recess 342 side prevents spheroids from is not discharged.
  • the cells (spheroids) that have been perfusion cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture with the surrounding internal space S2 communicated without being removed from the cell culture container A7. becomes possible.
  • the cell culture vessel A7 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 19 shows a cell culture container according to an eighth embodiment of the present disclosure.
  • the cell culture vessel A8 of the present embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second bag portion B2.
  • the second plate body 4 is not attached to the second film-like member 2 and is separate from the second film-like member. Further, the area of the second plate body 4 is made large, and the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the second bag portion B2 includes a second plate body 4 that is separate from the second film-like member 2 .
  • Materials suitable for culturing adherent cells include, for example, polycarbonate.
  • the material constituting the second plate member 4 is polycarbonate, there is a possibility that the adhesiveness between the second plate member 4 and the second film-like member 2 may be reduced.
  • the material of the second plate body 4 can be appropriately selected according to the purpose.
  • the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x.
  • cell culture can be appropriately continued on the large-area second plate body 4 even after shifting to expansion culture.
  • the cell culture vessel A8 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 20 shows a cell culture container according to the ninth embodiment of the present disclosure.
  • the cell culture vessel A9 of the present embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second plate body 4 .
  • the area of the second plate body 4 is increased.
  • the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x. According to such a configuration, cell culture can be appropriately continued on the large-area second plate body 4 even after shifting to expansion culture.
  • the cell culture vessel A9 of the present embodiment has the same effect as the above embodiment within the same range of configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 21 shows a cell culture container according to a tenth embodiment of the present disclosure.
  • the cell culture vessel A10 of the present embodiment further includes a third film member 9, and is appropriately modified accordingly.
  • the third film member 9 is positioned on the other side x2 in the first direction with respect to the second film member 2 (second bag portion B2).
  • the outer peripheral portion 92 of the third film member 9 is joined together with the outer peripheral portion 12 of the first film member 1 and the outer peripheral portion 22 of the second film member 2 by an appropriate means such as heat welding, and connected to each other.
  • a fourth space S4 is provided between the second bag portion B2 (the second film member 2 and the second plate member 4) on the other side x2 in the first direction (lower side in FIG. 21) and the third film member 9. formed.
  • one or a plurality of communication holes 24 are formed at appropriate locations near the outer peripheral portion 12. As shown in FIG.
  • the communication port 24 penetrates through the second film member 2 in the thickness direction, and the surrounding internal space S2 and the fourth space S4 are communicated through the communication port 24 . Also, when the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate body 4, the central internal space S1 and the peripheral internal space S2 communicate with each other. Thereby, when the suction pad 52 is not in close contact with the first bottom surface 41a, the central internal space S1 and the fourth space S4 communicate with each other through the communication port 24. As shown in FIG.
  • the communication port 24 described above corresponds to an example of "communication means".
  • a plurality of recesses 931 are formed on the bottom surface 93a of the third film member 9 facing the one side x1 in the first direction.
  • the plurality of recesses 931 are formed to include areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 93a.
  • the plurality of recesses 931 are densely arranged along the in-plane direction of the bottom surface 93a, like the plurality of recesses 411 shown in FIG.
  • the plurality of recesses 931 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the cell culture vessel A10 of the present embodiment includes a third film member 9 and has a fourth space S4 sandwiched between the third film member 9 and the first direction other side x2 of the second bag portion B2.
  • the central internal space S1 and the fourth space S4 communicate with each other through the communication port 24.
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate body 4 (preculture before expansion culture)
  • the first Adherent cells are cultured using the bottom surface 41a.
  • the cultured cells move to the fourth space S4 through the communication port 24 and reach the bottom surface 93a of the third film member 9. It is accommodated in the formed recess 931 .
  • cultured cells tend to aggregate three-dimensionally and form spheroids.
  • the cell culture vessel A10 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 22 shows a cell culture container according to the eleventh embodiment of the present disclosure.
  • the cell culture vessel A11 of the present embodiment has a port 99 provided on the third film member 9 instead of the communication port 24 provided on the second film member 2. It has been modified accordingly.
  • the third film member 9 is provided with a plurality of ports 99. Port holes 94 are formed at portions of the third film member 9 corresponding to the respective ports 99 .
  • Each port 19 has one side end (first direction other side x2 end) communicating with the outside, and the other side end (first direction one side x1 end) communicates with the fourth through port 99 of third film member 9 . It leads to space S42.
  • a tube 88 is connected to the port 19 of the first film member 1 and the port 99 of the third film member 9 . One end of tube 88 is connected to port 19 and the other end of tube 88 is connected to port 99 .
  • the communication port 24 is not formed. 22, an intermediate portion of the tube 88 is omitted.
  • the surrounding internal space S2 and the fourth space S4 are communicated via the tube 88. Also, when the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate body 4, the central internal space S1 and the peripheral internal space S2 communicate with each other. Thereby, the central internal space S1 and the fourth space S4 communicate with each other through the port 19, the tube 88 and the port 99 when the suction pad 52 is not in close contact with the first bottom surface 41a.
  • the port 19, tube 88 and port 99 described above correspond to an example of "communication means".
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11).
  • the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2.
  • the cell culture vessel A11 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, it is possible to shift to expansion culture during cell culture (perfusion culture) without changing the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the cell culture vessel A11 of the present embodiment includes a third film member 9 and has a fourth space S4 sandwiched between the third film member 9 and the first direction other side x2 of the second bag portion B2.
  • the suction pad 52 is not in close contact with the first bottom surface 41a (second bag portion B2) of the second plate member 4
  • the central internal space S1 and the fourth space S4 are connected through the port 19, the tube 88 and the port 99. communicate with.
  • the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate body 4 (preculture before expansion culture)
  • the first Adherent cells are cultured using the bottom surface 41a.
  • the cultured cells move to the fourth space S4 through the port 19, the tube 88 and the port 99, and are transferred to the third film member. 9 is housed in a recess 931 formed in the bottom surface 93a of .
  • cultured cells tend to aggregate three-dimensionally and form spheroids.
  • the cell culture vessel A11 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
  • FIG. 23 shows a cell culture vessel according to a twelfth embodiment of the present disclosure.
  • the cell culture vessel A12 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second bag portion B2.
  • the second plate body 4 is positioned on the other side x2 in the first direction with respect to the second film member 2 and has a substantially circular shape.
  • the second film member 2 does not have the second opening 21 and has a substantially circular shape.
  • the second plate member 4 and the second film member 2 overlap the central internal space S1 when viewed in the first direction x.
  • the second film-like member 2 is joined to the first bottom surface 41a (the surface facing the one side x1 in the first direction) of the second plate member 4 by, for example, heat sealing or adhesion.
  • the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other.
  • the first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 .
  • the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11).
  • the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate.
  • the cell culture vessel A12 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
  • the second plate body 4 is attached to the second film-like member 2 and positioned on the other side x2 in the first direction with respect to the second film-like member 2.
  • the bottom surface 23 a of the second film-like member 2 with which the suction pad 52 contacts can be maintained as a substantially flat surface corresponding to the first bottom surface 41 a of the second plate body 4 .
  • the suction pad 52 is brought into close contact with the bottom surface 23a, the close contact state of the suction pad 52 with the bottom surface 23a is appropriately maintained.
  • the region of the bottom surface 23a that overlaps the central internal space S1 when viewed in the first direction x can also be maintained as a substantially flat surface corresponding to the first bottom surface 41a of the second plate body 4 .
  • the bottom surface 23a can be maintained in a planar state suitable for cell culture.
  • the cell culture vessel A12 of the present embodiment has the same effect as the above embodiment within the same configuration range as the cell culture vessel A1 of the above embodiment.
  • the cell culture vessel of the present disclosure is not limited to the substantially circular configuration shown in each of the above embodiments, and may be, for example, substantially rectangular.
  • FIG. 24 shows a cell culture vessel A13 configured in a substantially rectangular shape. According to such a configuration, after the cells cultured in the first space S11 are formed into, for example, a sheet shape, the suction member 5 is released from the adhesion state of the suction pad 52 to the first bottom surface 41a, and the central inner space is removed. S1 (first space S11) and surrounding internal space S2 are communicated. Then, as shown in FIG. 25, the sheet-like cells Sc can be moved to one end (right end in the figure) of the cell culture container A13. After that, as shown in FIG.
  • a heat-sealed portion Hs is formed in the first film-like member 1 and the second film-like member 2 by heat-sealing such that the sheet-like cells Sc are included, and the heat-sealed portion Hs is formed. detach the As a result, the formed sheet-like cells Sc can be closedly packaged without being removed from the cell culture container A13. Therefore, the cell culture container A13 is convenient for transporting and storing the prepared sheet-like cells Sc without risk of contamination. Further, the cell culture container of the present disclosure may be configured such that a plurality of suction pads are in close contact with one second bag portion, and a plurality of central internal spaces corresponding to each suction pad are formed.

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Abstract

Provided is a container for cell culture use, which enables the transition to large-volume expansion culture without the need to transfer to another container so as to enable the continuation of cell perfusion culture.　The container is provided with: a first bag part that includes a first film-like member located on one side of a first direction and a first plate body attached to the first film-like member; a second bag part that includes a second film-like member located on the other side of the first direction relative to the first bag part and having an outer periphery connected to an outer periphery of the first film-like member; and an adsorption member attached to the first plate body. In the container, the adsorption member faces the other side of the first direction and has a ring-shaped adsorption pad capable of tightly adhering to the second bag part, and the first bag part has a first flow path and a second flow path each having such a structure that one side end is communicated with the outside and the other side end is positioned in a center internal space between the first bag part and the second bag part in the first direction in the adsorption member when observed in the first direction. The container is configured such that, when the adsorption pad is in a state where the adsorption pad is tightly adhered to the second bag part, the center internal space and a peripheral internal space surrounded by the first film-like member, the second film-like member and the adsorption member in the outside of the adsorption member when observed in the first direction are blocked from each other; while when the adsorption pad is in a state where the adsorption pad is not tightly adhered to the second bag part, the center internal space and the peripheral internal space are communicated with each other.

Description

細胞培養用容器cell culture vessel
　本開示は、細胞を培養するための細胞培養用容器に関し、特に、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に用いる細胞培養用容器に関する。 The present disclosure relates to a cell culture vessel for culturing cells, and in particular, a cell culture vessel used for perfusion culture that automatically supplies culture medium at a constant low speed and discharges the same amount of culture medium at the same time. Regarding.
　細胞培養においては、細胞の増殖に合わせ、新たな容器に細胞を移し替える作業を行う。その際大きな容器に移し替えたり容器の数を増やしたりする工程でコンタミネーションを起こしやすい。そのため、ディッシュやウェルプレート、培養フラスコ等、従来の培養容器による細胞培養では、コンタミネーションのリスクが大きかった。 In cell culture, the cells are transferred to a new container according to the growth of the cells. At that time, contamination is likely to occur in the process of transferring to a larger container or increasing the number of containers. Therefore, cell culture using conventional culture vessels such as dishes, well plates, and culture flasks has a high risk of contamination.
特開２０１４－１４３９９８号公報JP 2014-143998 A
　本開示は、このような事情のもとで考え出されたものであって、細胞を別の容器に移し替えること無く、容積を拡大した拡大培養に移行して細胞灌流培養を継続することが可能な細胞培養用容器を提供することを主たる課題とする。 The present disclosure has been conceived under such circumstances, and it is possible to continue cell perfusion culture by shifting to expansion culture with an expanded volume without transferring the cells to another container. The main object is to provide a container for cell culture that is capable of
　上記の課題を解決するため、本開示では、次の技術的手段を採用した。 In order to solve the above problems, the following technical measures are adopted in this disclosure.
　本開示によって提供される細胞培養用容器は、第１方向の一方側に位置する第１フィルム状部材、および前記第１フィルム状部材に取り付けられた第１プレート体、を含む第１バッグ部と、前記第１バッグ部に対して前記第１方向の他方側に位置し、外周部が前記第１フィルム状部材の外周部につながる第２フィルム状部材、を含む第２バッグ部と、前記第１プレート体に取り付けられた吸着部材と、を備え、前記吸着部材は、前記第１方向の他方側を向き、且つ前記第２バッグ部に密着可能な環状の吸着パッドを有し、前記第１バッグ部は、各々の一方側端が外部に通じ、且つ他方側端が、前記第１方向に見て前記吸着部材の内側で前記第１方向において前記第１バッグ部と前記第２バッグ部との間の中央内部空間に位置する第１流路および第２流路を有し、前記吸着パッドが前記第２バッグ部に密着する状態にあるとき、前記中央内部空間と、前記第１方向に見て前記吸着部材の外側で前記第１フィルム状部材、前記第２フィルム状部材および前記吸着部材で囲まれた周囲内部空間とが遮断され、前記吸着パッドが前記第２バッグ部に密着しない状態にあるとき、前記中央内部空間と前記周囲内部空間とが連通する。 A cell culture vessel provided by the present disclosure includes a first bag portion including a first film-like member positioned on one side in a first direction, and a first plate body attached to the first film-like member; a second film-like member located on the other side in the first direction with respect to the first bag portion and having an outer peripheral portion connected to the outer peripheral portion of the first film-like member; a suction member attached to one plate body, the suction member facing the other side in the first direction and having an annular suction pad capable of being in close contact with the second bag portion; One side end of each bag portion communicates with the outside, and the other side end is inside the adsorption member when viewed in the first direction, and is connected to the first bag portion and the second bag portion in the first direction. It has a first flow path and a second flow path located in the central internal space between the central internal space and the first direction when the suction pad is in close contact with the second bag portion. A state in which the suction pad does not come into close contact with the second bag portion because the inner space surrounded by the first film-like member, the second film-like member, and the suction member is cut off from the outside of the suction member. , the central interior space and the peripheral interior space are in communication.
　好ましい実施の形態においては、前記第１フィルム状部材は、内周側に第１開口を有する環状とされており、前記第１プレート体は、前記第１開口を塞いでいる。 In a preferred embodiment, the first film-like member has an annular shape with a first opening on the inner peripheral side, and the first plate body closes the first opening.
　好ましい実施の形態においては、前記第２バッグ部は、前記第２フィルム状部材とは別体の第２プレート体を含み、前記第２プレート体は、前記第１方向の一方側を向き、且つ略平坦な第１底面を有し、前記吸着パッドは、前記第１底面に密着可能である。 In a preferred embodiment, the second bag portion includes a second plate body separate from the second film-like member, the second plate body faces one side in the first direction, and It has a substantially flat first bottom surface, and the suction pad can be in close contact with the first bottom surface.
　好ましい実施の形態においては、前記第２バッグ部は、前記第２フィルム状部材に取り付けられた第２プレート体を含み、前記第２プレート体は、前記第１方向の一方側を向き、且つ略平坦な第１底面を有し、前記吸着パッドは、前記第１底面に密着可能である。 In a preferred embodiment, the second bag part includes a second plate body attached to the second film-like member, the second plate body faces one side in the first direction, and substantially It has a flat first bottom surface, and the suction pad can be brought into close contact with the first bottom surface.
　好ましい実施の形態においては、前記第２フィルム状部材は、内周側に第２開口を有する環状とされており、前記第２プレート体は、前記第２開口を塞いでいる。 In a preferred embodiment, the second film-like member has an annular shape with a second opening on the inner peripheral side, and the second plate body closes the second opening.
　好ましい実施の形態においては、前記第２バッグ部は、前記第２フィルム状部材に取り付けられた第２プレート体を含み、前記第２プレート体は、前記第２フィルム状部材に対して前記第１方向の他方側に位置し、且つ前記第１方向に見て前記中央内部空間と重なる。 In a preferred embodiment, said second bag part comprises a second plate body attached to said second film-like member, said second plate body being attached to said second film-like member by said first film-like member. located on the other side of the direction and overlapping the central interior space when viewed in the first direction.
　好ましい実施の形態においては、前記第１方向に見て、前記第２プレート体の面積は、前記第１プレート体よりも大きい。 In a preferred embodiment, the area of the second plate body is larger than that of the first plate body when viewed in the first direction.
　好ましい実施の形態においては、前記吸着部材は、前記第１プレート体を前記第１方向に貫通する少なくとも１つの吸引路を有し、前記吸引路は、一方側端が外部に通じており、且つ他方側端が前記吸着パッドの内側に通じる。 In a preferred embodiment, the adsorption member has at least one suction path that penetrates the first plate body in the first direction, one side end of the suction path communicates with the outside, and The other end communicates with the inside of the suction pad.
　好ましい実施の形態においては、前記第１フィルム状部材には、各々の一方側端が外部に通じ、且つ他方側端が前記周囲内部空間に通じる複数のポートが設けられている。 In a preferred embodiment, the first film-like member is provided with a plurality of ports each of which has one side end communicating with the outside and the other side end communicating with the surrounding internal space.
　好ましい実施の形態においては、前記第１プレート体は、前記第１方向に見て環状をなす第１環状板部と、前記第１方向に見て前記第１環状板部の内側を塞ぎ、且つ前記第１方向の一方側に張り出す膨出部と、を有し、前記第１環状板部には、ガス透過膜が取り付けられており、前記中央内部空間は、前記第２バッグ部と前記ガス透過膜および前記第１環状板部とに挟まれた第１空間と、前記ガス透過膜および前記膨出部に囲まれた第２空間と、を含み、前記第１プレート体は、各々の一方側端が外部に通じ、且つ他方側端が前記第２空間に通じる複数の第３流路を有する。 In a preferred embodiment, the first plate body includes a first annular plate portion having an annular shape when viewed in the first direction, and closing an inner side of the first annular plate portion when viewed in the first direction, and a bulging portion projecting to one side in the first direction, a gas permeable membrane is attached to the first annular plate portion, and the central internal space is formed between the second bag portion and the The first plate body includes a first space sandwiched between the gas permeable membrane and the first annular plate portion, and a second space surrounded by the gas permeable membrane and the bulging portion. It has a plurality of third flow paths with one side end communicating with the outside and the other side end communicating with the second space.
　好ましい実施の形態においては、前記第１プレート体には、液体が通過可能な膜フィルターが設けられており、前記中央内部空間は、前記膜フィルターに対して、前記第１方向の他方側に位置する前記第１空間および前記第１方向の一方側に位置する第３空間を含み、前記第１流路の他方側端は、前記第１空間に通じており、前記第２流路の他方側端は、前記第３空間に通じている。 In a preferred embodiment, the first plate body is provided with a liquid permeable membrane filter, and the central inner space is located on the other side in the first direction with respect to the membrane filter. and a third space located on one side in the first direction, the other side end of the first flow path communicates with the first space, and the other side of the second flow path The end communicates with the third space.
　好ましい実施の形態においては、前記第１プレート体のうち前記第３空間を形成する部分には、前記第１方向の一方側に凹む天井凹部が形成されており、前記天井凹部は、前記第２流路につながっている。 In a preferred embodiment, a portion of the first plate body that forms the third space is formed with a ceiling recess that is recessed toward one side in the first direction, and the ceiling recess is formed in the second space. connected to the stream.
　好ましい実施の形態においては、前記第２バッグ部において、前記第１方向の一方側を向く面のうち前記第１方向に見て前記吸着パッドの内側を含む領域には、稠密に配列された複数の凹部が設けられている。 In a preferred embodiment, in the second bag portion, a plurality of densely arranged suction pads are provided in a region of the surface facing one side in the first direction that includes the inner side of the suction pad when viewed in the first direction. is provided.
　好ましい実施の形態においては、外周部が前記第２フィルム状部材の前記外周部につながる第３フィルム状部材をさらに備え、前記第２バッグ部の前記第１方向の他方側と前記第３フィルム状部材とに挟まれた第４空間を有し、前記吸着パッドが前記第２バッグ部に密着しない状態にあるとき、前記中央内部空間と前記第４空間とが連通する連通手段を備える。 In a preferred embodiment, a third film-like member having an outer peripheral portion connected to the outer peripheral portion of the second film-like member is further provided, and the other side of the second bag portion in the first direction is connected to the third film-like member. and a communication means for communicating between the central internal space and the fourth space when the suction pad is not in close contact with the second bag portion.
　本開示のその他の特徴および利点は、添付図面を参照して以下に行う詳細な説明によって、より明らかとなろう。 Other features and advantages of the present disclosure will become clearer from the detailed description given below with reference to the accompanying drawings.
本開示の第１実施形態に係る細胞培養用容器を示す斜視図である。1 is a perspective view showing a cell culture container according to a first embodiment of the present disclosure; FIG. 図１に示す細胞培養用容器の平面図である。FIG. 2 is a plan view of the cell culture vessel shown in FIG. 1; 図２のＩＩＩ－ＩＩＩ線に沿う断面図である。3 is a cross-sectional view taken along line III-III of FIG. 2; FIG. 図２のＩＶ－ＩＶ線に沿う断面図である。3 is a cross-sectional view taken along line IV-IV of FIG. 2; FIG. 図２のＶ－Ｖ線に沿う断面図である。FIG. 3 is a cross-sectional view taken along line VV of FIG. 2; 本開示の第１実施形態に係る細胞培養用容器の使用状態を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure; 本開示の第１実施形態に係る細胞培養用容器の使用状態を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure; 本開示の第１実施形態に係る細胞培養用容器の使用状態を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure; 本開示の第１実施形態に係る細胞培養用容器の使用状態を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing the usage state of the cell culture vessel according to the first embodiment of the present disclosure; 本開示の第２実施形態に係る細胞培養用容器を示す斜視図である。Fig. 10 is a perspective view showing a cell culture container according to a second embodiment of the present disclosure; 図１０に示す培養用容器の平面図である。FIG. 11 is a plan view of the culture vessel shown in FIG. 10; 図１１のＸＩＩ－ＸＩＩ線に沿う断面図である。12 is a cross-sectional view along line XII-XII of FIG. 11; FIG. 本開示の第３実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a third embodiment of the present disclosure; 第２プレート体の第１底面に形成された複数の凹部の配列状態を示し、第１方向に見た図である。FIG. 10 is a diagram showing an arrangement state of a plurality of recesses formed on the first bottom surface of the second plate body, viewed in the first direction; 本開示の第４実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a fourth embodiment of the present disclosure; 本開示の第５実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a fifth embodiment of the present disclosure; 本開示の第６実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a sixth embodiment of the present disclosure; 本開示の第７実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a seventh embodiment of the present disclosure; 本開示の第８実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to an eighth embodiment of the present disclosure; 本開示の第９実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 4 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a ninth embodiment of the present disclosure; 本開示の第１０実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 10 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to a tenth embodiment of the present disclosure; 本開示の第１１実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 11 is a cross-sectional view, similar to FIG. 3, showing a cell culture vessel according to an eleventh embodiment of the present disclosure; 本開示の第１２実施形態に係る細胞培養用容器を示す、図３と同様の断面図である。FIG. 12 is a cross-sectional view similar to FIG. 3 showing a cell culture vessel according to a twelfth embodiment of the present disclosure; 本開示に係る細胞培養用容器の変形例を示す平面図である。FIG. 4 is a plan view showing a modification of the cell culture vessel according to the present disclosure; 図２４に示した細胞培養用容器の使用例を示す平面図である。FIG. 25 is a plan view showing a usage example of the cell culture vessel shown in FIG. 24; 図２４に示した細胞培養用容器の使用例を示す平面図である。FIG. 25 is a plan view showing a usage example of the cell culture vessel shown in FIG. 24;
　以下、本開示の好ましい実施の形態を図面を参照しつつ具体的に説明する。 Hereinafter, preferred embodiments of the present disclosure will be specifically described with reference to the drawings.
　本開示における「第１」、「第２」、「第３」等の用語は、単にラベルとして用いたものであり、必ずしもそれらの対象物に順列を付することを意図していない。 The terms "first", "second", "third", etc. in the present disclosure are merely used as labels and are not necessarily intended to give permutations to those objects.
＜第１実施形態＞
　図１～図３は、本開示の第１実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ１は、第１フィルム状部材１、第２フィルム状部材２、第１プレート体３、第２プレート体４、吸着部材５、ガス透過膜６、膜フィルター７を備えている。細胞培養用容器Ａ１は、継代培養から大容量の容器で行う拡大培養まで細胞培養を継続して行うのに用いられる。図１は、細胞培養用容器Ａ１の斜視図である。図２は、細胞培養用容器Ａ１の平面図である。 <First embodiment>
1 to 3 show a cell culture vessel according to a first embodiment of the present disclosure. The cell culture vessel A1 of the present embodiment comprises a first film member 1, a second film member 2, a first plate member 3, a second plate member 4, an adsorption member 5, a gas permeable membrane 6, and a membrane filter 7. I have. The cell culture vessel A1 is used for continuous cell culture from subculture to expansion culture performed in a large-capacity vessel. FIG. 1 is a perspective view of a cell culture vessel A1. FIG. 2 is a plan view of the cell culture vessel A1.
　図３～図５に示すように、第１フィルム状部材１および第２フィルム状部材２は、それぞれが環状（本実施形態では円環状）をなし、第１方向ｘに見て互いに重なっている。第１フィルム状部材１および第２フィルム状部材２は、柔軟性を有する薄膜状とされており、外力に追従して容易に撓み変形するものである。第１フィルム状部材１は、第１方向ｘの一方側（以下、適宜「第１方向一方側ｘ１」という。）に位置し、第２フィルム状部材２は、第１方向ｘの他方側（以下、適宜「第１方向他方側ｘ２」という。）に位置する。第１フィルム状部材１は内周側に第１開口１１を有し、第２フィルム状部材２は内周側に第２開口２１を有する。第１フィルム状部材１の外周部１２および第２フィルム状部材２の外周部２２は、例えば熱溶着等の適宜手段によって接合され、互いにつながっている。第１フィルム状部材１および第２フィルム状部材２の構成材料は特に限定されず、例えばポリエチレン、ポリオレフィン、スチレン系エラストマー、熱可塑性エラストマーなどが挙げられる。なお、第１フィルム状部材１および第２フィルム状部材２は、上記のように熱溶着で外周部１２，２２が接合された構成に限定されず、例えば外周部どうしが互いにつながるように一体的に形成された成形品であってもよい。 As shown in FIGS. 3 to 5, the first film-like member 1 and the second film-like member 2 each form an annular shape (in this embodiment, an annular shape) and overlap each other when viewed in the first direction x. . The first film-like member 1 and the second film-like member 2 are in the form of thin films having flexibility, and are easily flexurally deformed following an external force. The first film-like member 1 is positioned on one side in the first direction x (hereinafter referred to as “first direction one side x1”), and the second film-like member 2 is positioned on the other side in the first direction x ( hereinafter referred to as “first direction other side x2”). The first film-like member 1 has a first opening 11 on the inner peripheral side, and the second film-like member 2 has a second opening 21 on the inner peripheral side. The outer peripheral portion 12 of the first film-like member 1 and the outer peripheral portion 22 of the second film-like member 2 are joined by an appropriate means such as thermal welding and connected to each other. The constituent material of the first film-like member 1 and the second film-like member 2 is not particularly limited, and examples thereof include polyethylene, polyolefin, styrene-based elastomer, and thermoplastic elastomer. In addition, the first film-like member 1 and the second film-like member 2 are not limited to the structure in which the outer peripheral portions 12 and 22 are joined by heat welding as described above. It may be a molded product formed in the
　第１フィルム状部材１の適所には、複数のポート１９が設けられている。図３、図４に示すように、第１フィルム状部材１において各ポート１９に対応する箇所には、ポート用孔１３が形成されている。複数のポート１９については後述する。 A plurality of ports 19 are provided at appropriate locations on the first film member 1 . As shown in FIGS. 3 and 4 , port holes 13 are formed in the first film-like member 1 at locations corresponding to the respective ports 19 . The multiple ports 19 will be described later.
　図３～図５に示すように、第１プレート体３は、第２プレート体４に対して第１方向一方側ｘ１に位置する。第１プレート体３は、第１フィルム状部材１の第１開口１１を塞いでいる。第２プレート体４は、第１プレート体３に対して第１方向他方側ｘ２に位置する。第２プレート体４は、第２フィルム状部材２の第２開口２１を塞いでいる。 As shown in FIGS. 3 to 5, the first plate body 3 is located on the first direction one side x1 with respect to the second plate body 4. As shown in FIGS. The first plate body 3 closes the first opening 11 of the first film member 1 . The second plate body 4 is positioned on the other side x2 in the first direction with respect to the first plate body 3 . The second plate body 4 closes the second opening 21 of the second film member 2 .
　第１プレート体３は、第１環状板部３１、第２環状板部３２、膨出部３３およびフィルター押え３４を含んで構成される。第１環状板部３１は、第１方向ｘに見て環状をなしており、本実施形態では略円環状の板状部分を含んで構成される。第１環状板部３１は、第１流路３１１および吸着部材用凹溝３１２を有する。吸着部材用凹溝３１２は、第１方向他方側ｘ２に延びる環状筒部に囲まれた環状溝であり、吸着部材５を配置するための部位である。第１流路３１１については後述する。 The first plate body 3 includes a first annular plate portion 31, a second annular plate portion 32, a bulging portion 33 and a filter retainer . The first annular plate portion 31 has an annular shape when viewed in the first direction x, and includes a substantially annular plate portion in the present embodiment. The first annular plate portion 31 has a first flow path 311 and an adsorption member groove 312 . The adsorption member concave groove 312 is an annular groove surrounded by an annular tubular portion extending in the first direction other side x2, and is a portion for disposing the adsorption member 5 . The first channel 311 will be described later.
　膨出部３３は、第１環状板部３１の内周縁につながっている。膨出部３３は、第１方向ｘに見て第１環状板部３１の内側を塞いでいる。また、膨出部３３は、第１環状板部３１に対して第１方向一方側ｘ１に張り出している。膨出部３３は、複数の第３流路３３１を有する。第３流路３３１については後述する。 The bulging portion 33 is connected to the inner peripheral edge of the first annular plate portion 31 . The bulging portion 33 closes the inside of the first annular plate portion 31 when viewed in the first direction x. In addition, the bulging portion 33 protrudes toward the first direction one side x1 with respect to the first annular plate portion 31 . The bulging portion 33 has a plurality of third flow paths 331 . The 3rd flow path 331 is mentioned later.
　第２環状板部３２は、第１環状板部３１の外周縁に段差部を介してつながっており、円環状の板状部分である。この第２環状板部３２の外周部の下面（第１方向他方側ｘ２を向く面）には、例えば超音波溶着や接着などの手法により、第１フィルム状部材１の内周部が接合されている。これにより、第１プレート体３は第２フィルム状部材２の第１開口１１を塞いでいる。 The second annular plate portion 32 is connected to the outer peripheral edge of the first annular plate portion 31 via a stepped portion, and is an annular plate-like portion. The inner peripheral portion of the first film member 1 is joined to the lower surface of the outer peripheral portion of the second annular plate portion 32 (the surface facing the other side x2 in the first direction) by, for example, ultrasonic welding or adhesion. ing. Thereby, the first plate body 3 closes the first opening 11 of the second film member 2 .
　第１環状板部３１、第２環状板部３２および膨出部３３は、硬質素材からなり、例えば樹脂成形品である。第１環状板部３１、第２環状板部３２および膨出部３３の構成材料は特に限定されず、例えばポリスチレン、ポリカーボネート、ＣＯＰ（シクロオレフィンポリマー）などが挙げられる。 The first annular plate portion 31, the second annular plate portion 32, and the bulging portion 33 are made of a hard material, such as a resin molded product. The constituent materials of the first annular plate portion 31, the second annular plate portion 32, and the bulging portion 33 are not particularly limited, and examples thereof include polystyrene, polycarbonate, and COP (cycloolefin polymer).
　図３に示すように、フィルター押え３４は、膜フィルター７を挟む部材であり、第１環状板部３１の円筒部に嵌入されている。フィルター押え３４は、筒状部３４１、天井凹部３４２および第２流路３４３を有する。筒状部３４１は、円筒状であり、当該筒状部３４１が第１環状板部３１の円筒部に嵌入されている。天井凹部３４２は、筒状部３４１につながり、筒状部３４１の内側を塞いでいる。天井凹部３４２は、第１方向一方側ｘ１に凹んでいる。第１環状板部３１およびフィルター押え３４（筒状部３４１）は、膜フィルター７を第１方向ｘの両側から挟んでいる。詳細な図示説明は省略するが、筒状部３４１と第１環状板部３１の円筒部との間には、Ｏリングが介在している。これにより、第１環状板部３１とフィルター押え３４との嵌合部は密閉されている。第２流路３４３については後述する。 As shown in FIG. 3 , the filter presser 34 is a member that sandwiches the membrane filter 7 and is fitted into the cylindrical portion of the first annular plate portion 31 . The filter retainer 34 has a cylindrical portion 341 , a recessed ceiling portion 342 and a second flow path 343 . The cylindrical portion 341 has a cylindrical shape and is fitted into the cylindrical portion of the first annular plate portion 31 . The ceiling concave portion 342 is connected to the tubular portion 341 and closes the inside of the tubular portion 341 . The ceiling recessed portion 342 is recessed in the first direction one side x1. The first annular plate portion 31 and the filter retainer 34 (cylindrical portion 341) sandwich the membrane filter 7 from both sides in the first direction x. Although detailed illustration is omitted, an O-ring is interposed between the tubular portion 341 and the cylindrical portion of the first annular plate portion 31 . As a result, the fitting portion between the first annular plate portion 31 and the filter retainer 34 is sealed. The second flow path 343 will be described later.
　フィルター押え３４、硬質素材からなり、例えば樹脂成形品である。フィルター押え３４の構成材料は特に限定されず、例えばポリスチレン、ポリカーボネート、ＣＯＰなどが挙げられる。 The filter retainer 34 is made of a hard material, such as a resin molded product. The constituent material of the filter retainer 34 is not particularly limited, and examples thereof include polystyrene, polycarbonate, COP, and the like.
　上記の第１フィルム状部材１および第１プレート体３はそれぞれ、「第１バッグ部」の構成要素に相当する。なお、図面において、第１バッグ部の構成要素（第１フィルム状部材１および第１プレート体３）には、適宜符号Ｂ１を付している。 The first film-like member 1 and the first plate body 3 described above respectively correspond to constituent elements of the "first bag portion". In the drawings, the constituent elements of the first bag portion (the first film-like member 1 and the first plate body 3) are appropriately denoted by B1.
　第２プレート体４は、略円板状とされており、主板部４１を備えている。主板部４１は、平板状であり、第１底面４１ａを有する。第１底面４１ａは、第１方向一方側ｘ１を向き、略平坦である。第１底面４１ａが「略平坦」とは、後述の吸着部材５の吸着パッド５２が密着可能な平坦度を有すればよく、湾曲状など吸着パッド５２が密着困難な形状は含まないことを意味する。本実施形態では、第１底面４１ａは平滑な平面である。 The second plate body 4 has a substantially disc shape and includes a main plate portion 41 . The main plate portion 41 is flat and has a first bottom surface 41a. The first bottom surface 41a faces the first direction one side x1 and is substantially flat. The first bottom surface 41a being "substantially flat" means that the flatness is enough to allow the suction pads 52 of the suction member 5 described later to be in close contact, and does not include a shape such as a curved shape that makes it difficult for the suction pads 52 to be in close contact. do. In this embodiment, the first bottom surface 41a is a smooth plane.
　主板部４１（第２プレート体４）の外周部の上面（第１方向一方側ｘ１を向く第１底面４１ａ）には、例えばヒートシールや接着などの手法により、第２フィルム状部材２の内周部が接合されている。これにより、第２プレート体４は第２フィルム状部材２の第２開口２１を塞いでいる。第２プレート体４は、硬質素材からなり、例えば樹脂成形品である。第２プレート体４の構成材料は特に限定されず、例えばポリスチレン、ポリカーボネート、ＣＯＰなどが挙げられる。なお、図示した例では、第２プレート体４の外周部の上面（第１方向一方側ｘ１を向く第１底面４１ａ）に第２フィルム状部材２の内周部が接合されているが、第２プレート体４の外周部の下面（第１方向他方側ｘ２を向く面）に第２フィルム状部材２の内周部が接合されてもよい。 On the upper surface (the first bottom surface 41a facing the one side x1 in the first direction) of the outer periphery of the main plate portion 41 (the second plate body 4), the inner portion of the second film-like member 2 is applied, for example, by a technique such as heat sealing or adhesion. The perimeter is joined. Thereby, the second plate body 4 closes the second opening 21 of the second film member 2 . The second plate body 4 is made of a hard material, such as a resin molded product. The constituent material of the second plate body 4 is not particularly limited, and examples thereof include polystyrene, polycarbonate, and COP. In the illustrated example, the inner peripheral portion of the second film member 2 is joined to the upper surface of the outer peripheral portion of the second plate member 4 (the first bottom surface 41a facing the one side x1 in the first direction). The inner peripheral portion of the second film member 2 may be joined to the lower surface of the outer peripheral portion of the two-plate body 4 (the surface facing the other side x2 in the first direction).
　上記の第２フィルム状部材２および第２プレート体４はそれぞれ、「第２バッグ部」の構成要素に相当する。なお、図面において、第２バッグ部の構成要素（第２フィルム状部材２および第２プレート体４）には、適宜符号Ｂ２を付している。 The second film-like member 2 and the second plate body 4 described above each correspond to constituent elements of the "second bag portion". In the drawings, the constituent elements of the second bag portion (the second film-like member 2 and the second plate body 4) are appropriately denoted by B2.
　図３～図５に示した吸着部材５は、細胞培養用容器Ａ１の内部空間を中央内部空間Ｓ１および周囲内部空間Ｓ２に遮断するためのものである。吸着部材５は、第１方向ｘに見て全体として環状をなす。吸着部材５は、第１プレート体３に取り付けられている。吸着部材５は、ブロック部５１、吸着パッド５２および吸引ポート５３を有する。ブロック部５１は、その大部分が吸着部材用凹溝３１２に収容されている。ブロック部５１は、吸着部材用凹溝３１２に圧入されており、必要に応じて接着等により固定される。吸着パッド５２は、ブロック部５１の第１方向他方側ｘ２につながり、第１方向他方側ｘ２を向く環状の吸盤である。より具体的には、吸着パッド５２は、ブロック部５１の下端（第１方向他方側ｘ２端）から径方向外方および径方向内方の双方に延びる一対の環状リップ部を有する。この吸着パッド５２は、図６に示すように、第２プレート体４の第１底面４１ａに密着可能である。 The adsorption member 5 shown in FIGS. 3 to 5 is for isolating the internal space of the cell culture container A1 into the central internal space S1 and the peripheral internal space S2. The adsorption member 5 has an annular shape as a whole when viewed in the first direction x. The adsorption member 5 is attached to the first plate body 3 . The adsorption member 5 has a block portion 51 , an adsorption pad 52 and a suction port 53 . A large portion of the block portion 51 is accommodated in the suction member recessed groove 312 . The block portion 51 is press-fitted into the suction member recessed groove 312 and fixed by adhesion or the like as necessary. The suction pad 52 is an annular suction cup connected to the first direction other side x2 of the block portion 51 and facing the first direction other side x2. More specifically, the suction pad 52 has a pair of annular lip portions extending both radially outward and radially inward from the lower end of the block portion 51 (the other side x2 end in the first direction). The suction pad 52 can be brought into close contact with the first bottom surface 41a of the second plate body 4, as shown in FIG.
　図６に示すように、吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態にあるとき、中央内部空間Ｓ１と周囲内部空間Ｓ２とが遮断される。中央内部空間Ｓ１は、第１方向ｘに見て吸着部材５の内側で、第１方向ｘにおいて第１フィルム状部材１（第１バッグ部Ｂ１）および第２フィルム状部材２（第２バッグ部Ｂ２）の間に位置する。周囲内部空間Ｓ２は、第１方向ｘに見て吸着部材５の外側に位置し、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれている。その一方、例えば吸着パッド５２が第２プレート体４の第１底面４１ａから離れ、当該吸着パッド５２が第１底面４１ａに密着しない状態にあるとき、中央内部空間Ｓ１と周囲内部空間Ｓ２とが連通する。 As shown in FIG. 6, when the suction pad 52 is in close contact with the first bottom surface 41a of the second plate body 4, the central internal space S1 and the surrounding internal space S2 are cut off. The central internal space S1 is the inside of the adsorption member 5 when viewed in the first direction x, and the first film-like member 1 (first bag portion B1) and the second film-like member 2 (second bag portion) in the first direction x. B2). The surrounding internal space S<b>2 is located outside the adsorption member 5 when viewed in the first direction x and is surrounded by the first film member 1 , the second film member 2 and the adsorption member 5 . On the other hand, for example, when the suction pad 52 is separated from the first bottom surface 41a of the second plate body 4 and the suction pad 52 is not in close contact with the first bottom surface 41a, the central internal space S1 and the peripheral internal space S2 communicate with each other. do.
　図４に示すように、吸引ポート５３は、ポンプなどの吸引手段（図示略）を接続し、吸着パッド５２の内側を吸引するためのものである。吸引ポート５３は、延出部５３１、吸引路５３２および突起部５３３を有する。延出部５３１は、ブロック部５１から第１方向一方側ｘ１に延びる円筒状とされており、横断面円形の外周面を有する。延出部５３１は、第１環状板部３１に形成された貫通孔３１３を通じて当該第１環状板部３１よりも第１方向一方側ｘ１に突出している。本実施形態において、延出部５３１の先端（第１方向一方側ｘ１端）には、延出部５３１の外周面よりも外向きに突出する突起部５３３が設けられている。貫通孔３１３の内径寸法は、自然状態において延出部５３１の外径寸法よりも僅かに小さくされている。延出部５３１は、貫通孔３１３に圧入されている。これにより、貫通孔３１３（第１環状板部３１）は、延出部５３１の外周面に密着（当接）して当該外周面を囲っている。 As shown in FIG. 4, the suction port 53 is for connecting suction means (not shown) such as a pump to suck the inside of the suction pad 52 . The suction port 53 has an extension 531 , a suction path 532 and a protrusion 533 . The extending portion 531 has a cylindrical shape extending from the block portion 51 in the first direction one side x1 and has an outer peripheral surface with a circular cross section. The extending portion 531 protrudes from the first annular plate portion 31 to the first direction one side x1 through the through hole 313 formed in the first annular plate portion 31 . In the present embodiment, a projecting portion 533 that protrudes outward from the outer peripheral surface of the extending portion 531 is provided at the tip of the extending portion 531 (first direction one side x1 end). The inner diameter dimension of the through hole 313 is slightly smaller than the outer diameter dimension of the extending portion 531 in the natural state. The extending portion 531 is press-fitted into the through hole 313 . As a result, the through hole 313 (the first annular plate portion 31) is in close contact with (contacts with) the outer peripheral surface of the extending portion 531 to surround the outer peripheral surface.
　また、本実施形態では、図４等から理解されるように、上下方向（第１方向）に見て、第１環状板部３１における貫通孔３１３の周囲は、延出部５３１の先端に設けられた突起部５３３と重なっている。突起部５３３は、第１環状板部３１の上方に隣接している。これにより、第１環状板部３１の上端（第１方向一方側ｘ１端）には、突起部５３３が当接し得る。 Further, in this embodiment, as can be understood from FIG. It overlaps with the protruding portion 533 that is formed. The projecting portion 533 is adjacent above the first annular plate portion 31 . Thereby, the protrusion 533 can come into contact with the upper end (first direction one side x1 end) of the first annular plate portion 31 .
　吸引路５３２は、第１プレート体３を第１方向ｘに貫通している。吸引路５３２は、一方側端（第１方向一方側ｘ１端）が外部に通じており、他方側端（第１方向他方側ｘ２端）が吸着パッド５２の内側に通じている。 The suction path 532 penetrates the first plate body 3 in the first direction x. One side end (first direction one side x1 end) of the suction path 532 communicates with the outside, and the other side end (first direction other side x2 end) communicates with the inside of the suction pad 52 .
　吸着部材５は、例えば弾性変形容易な軟質材料により構成される。吸着部材５の構成材料は特に限定されず、例えばシリコーンゴム、熱可塑性エラストマーなどが挙げられる。 The adsorption member 5 is made of, for example, a soft material that is easily elastically deformable. The constituent material of the adsorption member 5 is not particularly limited, and examples thereof include silicone rubber and thermoplastic elastomer.
　図３～図５に示したガス透過膜６は、ガス透過性を有する薄状膜である。その一方、ガス透過膜６は、液体は通過することができず、液非透過性である。ガス透過膜６は、第１プレート体３の第１環状板部３１に取り付けられている。具体的には、ガス透過膜６は、第１環状板部３１の下面（第１方向他方側ｘ２を向く面）に例えば接着等の適宜手段により密着状態で固定される。このガス透過膜６が設けられることで、中央内部空間Ｓ１は、互いに区画された第１空間Ｓ１１および第２空間Ｓ１２を含む。第１空間Ｓ１１は、第２プレート体４（第１底面４１ａ）とガス透過膜６および第１環状板部３１とに挟まれている。第２空間Ｓ１２は、ガス透過膜６および膨出部３３に囲まれている。ガス透過膜６の構成材料は特に限定されず、例えばＰＥＴ（ポリエステル）、あるいはＰＣ（ポリカーボネート）製の疎水性フィルターなどが挙げられる。 The gas permeable membrane 6 shown in FIGS. 3 to 5 is a thin membrane having gas permeability. On the other hand, the gas-permeable membrane 6 is impermeable to liquid and impermeable to liquid. The gas permeable membrane 6 is attached to the first annular plate portion 31 of the first plate body 3 . Specifically, the gas permeable film 6 is fixed in close contact with the lower surface of the first annular plate portion 31 (the surface facing the other side x2 in the first direction) by an appropriate means such as adhesion. By providing the gas permeable membrane 6, the central internal space S1 includes a first space S11 and a second space S12 that are separated from each other. The first space S<b>11 is sandwiched between the second plate body 4 (first bottom surface 41 a ), the gas permeable film 6 and the first annular plate portion 31 . The second space S<b>12 is surrounded by the gas permeable membrane 6 and the bulging portion 33 . The constituent material of the gas permeable membrane 6 is not particularly limited, and examples thereof include a hydrophobic filter made of PET (polyester) or PC (polycarbonate).
　図３に示した膜フィルター７は、液体が通過可能な液通過性の膜体である。膜フィルター７は、例えば培養液などの液体が通過可能である。また、空気などの気体も膜フィルター７を通過可能である。その一方、培養細胞などの液体以外の内容物は、膜フィルター７を通過することができずに捕捉される。この膜フィルター７が設けられることで、中央内部空間Ｓ１は、区画された第３空間Ｓ１３を含む。第３空間Ｓ１３は、膜フィルター７に対して第１方向一方側ｘ１に位置する。第３空間Ｓ１３は、膜フィルター７およびフィルター押え３４に囲まれている。上記の第１空間Ｓ１１は、膜フィルター７に対して第１方向他方側ｘ２に位置する。膜フィルター７の構成材料は特に限定されず、例えばＰＥＴ（ポリエステル）、あるいはＰＣ（ポリカーボネート）製の親水化フィルターなどが挙げられる。 The membrane filter 7 shown in FIG. 3 is a liquid-permeable membrane through which liquid can pass. The membrane filter 7 can pass a liquid such as a culture solution. Gases such as air can also pass through the membrane filter 7 . On the other hand, non-liquid contents such as cultured cells cannot pass through the membrane filter 7 and are captured. By providing the membrane filter 7, the central internal space S1 includes a partitioned third space S13. The third space S13 is positioned on the one side x1 in the first direction with respect to the membrane filter 7 . The third space S13 is surrounded by the membrane filter 7 and the filter retainer . The first space S11 is positioned on the other side x2 in the first direction with respect to the membrane filter 7 . The constituent material of the membrane filter 7 is not particularly limited, and examples thereof include hydrophilic filters made of PET (polyester) or PC (polycarbonate).
　第１流路３１１、第２流路３４３および複数の第３流路３３１は、外部と細胞培養用容器Ａ１の内部空間とに通じる筒状の連通路である。第１流路３１１および第２流路３４３は、各々、一方側端（第１方向一方側ｘ１端）が外部に通じ、他方側端（第１方向他方側ｘ２端）が中央内部空間Ｓ１に位置する。具体的には、図３～図５に示すように、第１流路３１１の他方側端（第１方向他方側ｘ２端）は、第１空間Ｓ１１に通じている。図３に示すように、第２流路３４３の他方側端（第１方向他方側ｘ２端）は、第３空間Ｓ１３に通じている。第３空間Ｓ１３を規定するフィルター押え３４の天井凹部３４２は、第２流路３４３につながっている。 The first flow path 311, the second flow path 343 and the plurality of third flow paths 331 are cylindrical communicating paths that communicate with the outside and the internal space of the cell culture vessel A1. Each of the first flow path 311 and the second flow path 343 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) to the central internal space S1. To position. Specifically, as shown in FIGS. 3 to 5, the other side end (first direction other side x2 end) of the first flow path 311 communicates with the first space S11. As shown in FIG. 3, the other side end (first direction other side x2 end) of the second flow path 343 communicates with the third space S13. A ceiling recessed portion 342 of the filter retainer 34 that defines the third space S13 is connected to a second flow path 343 .
　複数の第３流路３３１は、各々、一方側端（第１方向一方側ｘ１端）が外部に通じ、他方側端（第１方向他方側ｘ２端）が第２空間Ｓ１２に通じる。 Each of the plurality of third flow paths 331 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) communicated with the second space S12.
　図３、図４に示した複数のポート１９は、各々、その一方側端（第１方向一方側ｘ１端）が外部に通じており、他方側端（第１方向他方側ｘ２端）が第１フィルム状部材１のポート用孔１３を介して周囲内部空間Ｓ２に通じている。 Each of the plurality of ports 19 shown in FIGS. 3 and 4 has one side end (first direction one side x1 end) communicated with the outside, and the other side end (first direction other side x2 end) communicates with the outside. 1 It communicates with the surrounding internal space S2 through the port hole 13 of the film-like member 1 .
　次に、細胞培養用容器Ａ１の使用方法および作用について、図６～図９を参照しつつ説明する。 Next, the usage method and action of the cell culture vessel A1 will be described with reference to FIGS. 6 to 9.
　細胞培養用容器Ａ１は、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を容器外に排出する灌流培養に使用される。なお、細胞培養用容器Ａ１に収容される培養細胞や培養液については、特に限定されるものではない。 The cell culture vessel A1 is used for perfusion culture in which the culture medium is automatically supplied at a constant low speed at all times, and the same amount of culture medium is discharged outside the vessel at the same time. In addition, the cultured cells and the culture solution to be accommodated in the cell culture vessel A1 are not particularly limited.
　細胞培養用容器Ａ１を用いた細胞培養では、図６に示すように、あらかじめ吸着部材５の吸着パッド５２を第２プレート体４の第１底面４１ａに密着させておく。第１底面４１ａは平滑で接着細胞の培養面としても好適に使用できる。ここで、第１プレート体３を第１方向他方側ｘ２に押し付けることで、第１プレート体３に取り付けられた吸着部材５の吸着パッド５２を第１底面４１ａに密着させることができる。このとき、上述のように、中央内部空間Ｓ１と周囲内部空間Ｓ２とが遮断される。また、図４に示した吸引ポート５３（吸引路５３２）に接続された吸引手段（図示略）によって吸着パッド５２の内側が吸引され、吸着パッド５２（吸着部材５）による第１底面４１ａの吸着力が高められる。これにより、中央内部空間Ｓ１と周囲内部空間Ｓ２とは、より確実に遮断される。 In cell culture using the cell culture vessel A1, the adsorption pad 52 of the adsorption member 5 is brought into close contact with the first bottom surface 41a of the second plate body 4 in advance, as shown in FIG. The first bottom surface 41a is smooth and can be suitably used as a culture surface for adherent cells. Here, by pressing the first plate body 3 against the first direction other side x2, the suction pad 52 of the suction member 5 attached to the first plate body 3 can be brought into close contact with the first bottom surface 41a. At this time, as described above, the central internal space S1 and the surrounding internal space S2 are cut off. Further, the inside of the suction pad 52 is sucked by suction means (not shown) connected to the suction port 53 (suction path 532) shown in FIG. power is increased. As a result, the central internal space S1 and the surrounding internal space S2 are more reliably cut off.
　次に、第１流路３１１および第２流路３４３を介して細胞培養用容器Ａ１に培養液の供給および空気の排出を行い、中央内部空間Ｓ１を培養液で満たす。ここで、第１流路３１１および第２流路３４３にはそれぞれチューブ８１が接続されており、第１流路３１１の一方側端（第１方向一方側ｘ１端）には、チューブ８１を介して灌流用ポンプ（図示略）が接続され、第２流路３４３の一方側端（第１方向一方側ｘ１端）には、チューブ８１を介して廃液用容器（図示略）が接続される。なお、中央内部空間Ｓ１を培養液で満たす際、培養液とともに培養細胞を中央内部空間Ｓ１に供給する。 Next, the culture solution is supplied to the cell culture vessel A1 and the air is discharged through the first channel 311 and the second channel 343 to fill the central internal space S1 with the culture solution. Here, a tube 81 is connected to each of the first flow path 311 and the second flow path 343 , and one end of the first flow path 311 (first direction one side x1 end) is connected via the tube 81 . A perfusion pump (not shown) is connected to the second channel 343 , and a waste liquid container (not shown) is connected to one end of the second flow path 343 (first direction one side x1 end) via a tube 81 . In addition, when filling the central internal space S1 with the culture solution, the cultured cells are supplied to the central internal space S1 together with the culture solution.
　第１流路３１１の他方側端（第１方向他方側ｘ２端）は、中央内部空間Ｓ１の第１空間Ｓ１１に通じている。これにより、第１空間Ｓ１１は、第１流路３１１を介して供給される培養液で満たされる。また、第１流路３１１を介して供給される培養細胞も第１空間Ｓ１１に流れ込む。第１空間Ｓ１１にある空気は、気体が通過可能な膜フィルター７を通過し、第３空間Ｓ１３を経て第２流路３４３から外部に排出される。なお、第１プレート体３に設けられた第１流路３１１および第２流路３４３の配置は特に限定されない。本実施形態では、第１流路３１１および第２流路３４３は、第１方向ｘに見て第１プレート体３の中央部を挟んで互いに反対側に離間して配置されている。このような構成によれば、中央内部空間Ｓ１（第１空間Ｓ１１）にムラなく培養液を還流可能となる。 The other side end (first direction other side x2 end) of the first flow path 311 communicates with the first space S11 of the central internal space S1. As a result, the first space S11 is filled with the culture medium supplied through the first channel 311 . Cultured cells supplied via the first channel 311 also flow into the first space S11. The air in the first space S11 passes through the gas-permeable membrane filter 7, passes through the third space S13, and is discharged from the second flow path 343 to the outside. The arrangement of the first flow path 311 and the second flow path 343 provided in the first plate body 3 is not particularly limited. In this embodiment, the first flow path 311 and the second flow path 343 are spaced apart from each other on opposite sides of the central portion of the first plate body 3 when viewed in the first direction x. According to such a configuration, the culture solution can be uniformly circulated to the central internal space S1 (first space S11).
　第１空間Ｓ１１の第１方向一方側ｘ１にある膜フィルター７は液体が通過可能である。このため、第１空間Ｓ１１を満たした培養液は、膜フィルター７を通過して第３空間Ｓ１３に流れ込み、第３空間Ｓ１３を満たした後に第２流路３４３を介して外部に排出される。図７は、第１空間Ｓ１１および第３空間Ｓ１３が培養液Ｃで満たされた状態を示す。 Liquid can pass through the membrane filter 7 located on the first direction one side x1 of the first space S11. Therefore, the culture solution filling the first space S11 passes through the membrane filter 7 and flows into the third space S13. 7 shows a state in which the first space S11 and the third space S13 are filled with the culture solution C. FIG.
　細胞培養用容器Ａ１の第１空間Ｓ１１に少量の空気が残存する場合がある。膜フィルター７の上側（第１方向一方側ｘ１）に位置する第３空間Ｓ１３を形成するフィルター押え３４は、第１方向一方側ｘ１に凹む天井凹部３４２が形成されており、当該天井凹部３４２は第２流路３４３につながっている。このような構成によれば、第２流路３４３が上位に位置するように細胞培養用容器Ａ１を傾けると、第１空間Ｓ１１内の気泡は、膜フィルター７を通過して第３空間Ｓ１３に移動する。この状態で灌流用ポンプを作動させることにより、第３空間Ｓ１３に残存する気泡は天井凹部３４２を通じて第２流路３４３内に移動し、細胞培養用容器Ａ１の外部に排出することができる。そして、灌流培養を行う際には、第１流路３１１を通じて新しい培養液Ｃが第１空間Ｓ１１（中央内部空間Ｓ１）に供給され、かつ第２流路３４３を通じて第３空間Ｓ１３（中央内部空間Ｓ１）の培養液Ｃが外部に排出される。灌流培養工程においては、細胞培養用容器Ａ１内（中央内部空間Ｓ１）の培養液Ｃを僅かずつ入れ替え続ける。 A small amount of air may remain in the first space S11 of the cell culture container A1. The filter presser 34 forming the third space S13 located above the membrane filter 7 (first direction one side x1) is formed with a ceiling recess 342 recessed in the first direction one side x1. It is connected to the second channel 343 . According to such a configuration, when the cell culture container A1 is tilted so that the second flow path 343 is positioned at the top, air bubbles in the first space S11 pass through the membrane filter 7 and enter the third space S13. Moving. By operating the perfusion pump in this state, air bubbles remaining in the third space S13 can move into the second channel 343 through the ceiling recess 342 and be discharged to the outside of the cell culture vessel A1. When perfusion culture is performed, a new culture solution C is supplied to the first space S11 (central internal space S1) through the first channel 311, and is supplied to the third space S13 (central internal space S1) through the second channel 343. The culture solution C in S1) is discharged to the outside. In the perfusion culture step, the culture medium C in the cell culture vessel A1 (the central internal space S1) is gradually replaced.
　灌流スピードは極めてゆっくりしたものなので、培養細胞が自重に逆らい浮遊して天井凹部３４２側に設けられた膜フィルター７に吸い寄せられることは殆どない。培養液Ｃ中にある培養細胞は、万一膜フィルター７に到達した場合でも膜フィルター７を通過することができない。これにより、第１空間Ｓ１１において培養細胞が培養液Ｃ中に浮遊しても、培養液Ｃ中の培養細胞が第２流路３４３から不当に排出されることは防止される。 Since the perfusion speed is extremely slow, the cultured cells float against their own weight and are almost never attracted to the membrane filter 7 provided on the side of the recess 342 on the ceiling. Cultured cells in the culture solution C cannot pass through the membrane filter 7 even if they reach the membrane filter 7 . As a result, even if the cultured cells float in the culture solution C in the first space S11, the cultured cells in the culture solution C are prevented from being unduly discharged from the second channel 343 .
　第１プレート体３の第１環状板部３１には、ガス透過膜６が取り付けられている。中央内部空間Ｓ１は、第２プレート体４の第１底面４１ａとガス透過膜６および第１環状板部３１とに挟まれた第１空間Ｓ１１と、ガス透過膜６および膨出部３３に囲まれた第２空間Ｓ１２と、を含む。膨出部３３（第１プレート体３）は、複数の第３流路３３１を有し、各第３流路３３１は、一方側端（第１方向一方側ｘ１端）が外部に通じ、他方側端（第１方向他方側ｘ２端）が第２空間Ｓ１２に通じる。このような構成によれば、必要に応じて、複数の第３流路３３１を通じて所定のガスを膨出部３３に供給し、且つ排出することで、第２空間Ｓ１２を所定のガス雰囲気に維持することができる。そして、第１空間Ｓ１１の内容物（培養液Ｃおよび培養細胞）については、ガス透過膜６を介して第２空間Ｓ１２との通気状態が維持され、第２空間Ｓ１２のガスを取り込むことが可能である。さらに、第１空間Ｓ１１にある内容物ガス雰囲気を、第３流路３３１に供給するガス種を変更することで第２空間Ｓ１２のガス雰囲気を、容易、且つ迅速に変更することができる。例えばインキュベータ等を用いて所定のガス雰囲気状態を作る場合と比べて、簡単な構成で所望のガス雰囲気の通気を行うことが可能である。 A gas permeable membrane 6 is attached to the first annular plate portion 31 of the first plate body 3 . The central internal space S1 is surrounded by a first space S11 sandwiched between the first bottom surface 41a of the second plate body 4, the gas permeable membrane 6 and the first annular plate portion 31, the gas permeable membrane 6 and the bulging portion 33. and a second space S12. The bulging portion 33 (first plate body 3) has a plurality of third flow paths 331, and one side end (first direction one side x1 end) of each third flow path 331 communicates with the outside. The side end (first direction other side x2 end) communicates with the second space S12. According to such a configuration, the second space S12 is maintained in a predetermined gas atmosphere by supplying a predetermined gas to the bulging portion 33 through the plurality of third flow paths 331 and discharging the predetermined gas as necessary. can do. The contents of the first space S11 (the culture medium C and the cultured cells) are maintained in a ventilated state with the second space S12 through the gas permeable membrane 6, and the gas in the second space S12 can be taken in. is. Furthermore, by changing the type of gas supplied to the third flow path 331, the gas atmosphere of the content gas in the first space S11 can be changed easily and quickly in the second space S12. For example, compared with the case of creating a predetermined gas atmosphere using an incubator or the like, it is possible to ventilate a desired gas atmosphere with a simple configuration.
　細胞培養用容器Ａ１を用いた灌流培養では、細胞培養の段階に応じて、培養液Ｃを収容する容器が大容量である拡大培養に移行する。本実施形態の細胞培養用容器Ａ１は、第１フィルム状部材１および第２フィルム状部材２を備える。第１フィルム状部材１および第２フィルム状部材２は、各々が環状で外周部１２，２２どうしが互いにつながり、内周側の第１開口１１および第２開口２１が第１プレート体３および第２プレート体４によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、培養液Ｃで満たされた中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。細胞培養用容器Ａ１への培養液Ｃの供給量を多くし、第１フィルム状部材１および第２フィルム状部材２が膨らんだ状態とすることで、培養液Ｃの収容空間（中央内部空間Ｓ１（第１空間Ｓ１１）および周囲内部空間Ｓ２）の容積が拡大されて大容量となる（図９参照）。これにより、大容量の拡大培養に移行することができる。本実施形態の細胞培養用容器Ａ１によれば、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the perfusion culture using the cell culture vessel A1, depending on the stage of cell culture, the process shifts to expansion culture in which the vessel containing the culture solution C has a large capacity. A cell culture vessel A1 of the present embodiment includes a first film-like member 1 and a second film-like member 2 . The first film-like member 1 and the second film-like member 2 are each annular, and the outer peripheral portions 12 and 22 are connected to each other. 2 It is blocked by the plate body 4 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , and the central internal space S1 (first space S11) filled with the culture medium C is cut off. By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. By increasing the supply amount of the culture solution C to the cell culture container A1 and setting the first film-like member 1 and the second film-like member 2 in a swollen state, the storage space for the culture solution C (the central internal space S1 The volumes of (the first space S11) and the surrounding internal space S2) are expanded to a large capacity (see FIG. 9). As a result, it is possible to shift to large-capacity expansion culture. According to the cell culture container A1 of the present embodiment, it is possible to shift to expansion culture in the middle of cell culture (perfusion culture) and continue cell culture (perfusion culture) without moving the container. be. Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態において、第１フィルム状部材１には、複数のポート１９が設けられている。各ポート１９は、一方側端（第１方向一方側ｘ１端）が外部に通じており、他方側端（第１方向他方側ｘ２端）が周囲内部空間Ｓ２に通じている。このような構成によれば、図９に示すように、例えば複数のポート１９それぞれにチューブ８２を接続し、一部のポート１９にはチューブ８２を介して溶液注入用ポンプ（図示略）を接続し、残りのポート１９にはチューブ８２を介して廃液排出用ポンプ、さらにチューブ８２を介して廃液用容器（図示略）を接続する。そうすることで、周囲内部空間Ｓ２へ培養液Ｃを効率よく供給することができ、迅速に拡大培養に移行することが可能である。図９に示すように、複数のポート１９のうち排出用に用いるものは、周囲内部空間Ｓ２に向かってチューブ８３を介して廃液排出口８４が接続されている。廃液排出口８４には、フィルター８５が設けられている（図９以外の図面においては、チューブ８３、廃液排出口８４およびフィルター８５を省略）。これにより、周囲内部空間Ｓ２において培養細胞が培養液Ｃ中に浮遊しても、培養液Ｃ中の培養細胞がポート１９から不当に排出されることは防止される。 In this embodiment, the first film-like member 1 is provided with a plurality of ports 19 . One side end (first direction one side x1 end) of each port 19 communicates with the outside, and the other side end (first direction other side x2 end) communicates with the surrounding internal space S2. According to such a configuration, as shown in FIG. 9, for example, a tube 82 is connected to each of the plurality of ports 19, and a solution injection pump (not shown) is connected to some of the ports 19 via the tube 82. The remaining port 19 is connected via a tube 82 to a waste liquid discharge pump and further via a tube 82 to a waste liquid container (not shown). By doing so, the culture medium C can be efficiently supplied to the surrounding internal space S2, and it is possible to quickly shift to expansion culture. As shown in FIG. 9, one of the plurality of ports 19 used for discharge is connected to a waste liquid discharge port 84 via a tube 83 toward the surrounding internal space S2. The waste liquid outlet 84 is provided with a filter 85 (the tube 83, the waste liquid outlet 84 and the filter 85 are omitted in the drawings other than FIG. 9). As a result, even if the cultured cells float in the culture solution C in the surrounding internal space S2, the cultured cells in the culture solution C are prevented from being improperly discharged from the port 19. FIG.
　中央内部空間Ｓ１と周囲内部空間Ｓ２とが連通した段階においては、培養液Ｃの供給量と排出量を適宜調節する。これにより、図９に示すように、中央内部空間Ｓ１および周囲内部空間Ｓ２が培養液Ｃで満たされることはなく、細胞培養用容器Ａ１内の培養液Ｃの液面の位置が調整される。したがって、中央内部空間Ｓ１および周囲内部空間Ｓ２のうち第１方向一方側ｘ１には、培養液Ｃが存在しない空間が存在する。なお、中央内部空間Ｓ１（第１空間Ｓ１１）のみで灌流培養をする段階におけるガス雰囲気調整は第３流路３３１を使用したが、周囲内部空間Ｓ２を連通させた段階におけるガス雰囲気調整は、第１流路３１１および第２流路３４３を使用して行うことが可能である。 At the stage when the central internal space S1 and the peripheral internal space S2 are in communication, the supply amount and discharge amount of the culture solution C are appropriately adjusted. As a result, as shown in FIG. 9, the central internal space S1 and the peripheral internal space S2 are not filled with the culture solution C, and the position of the liquid surface of the culture solution C in the cell culture vessel A1 is adjusted. Therefore, in the first direction one side x1 of the central internal space S1 and the peripheral internal space S2, there is a space where the culture solution C does not exist. Although the third channel 331 was used for the gas atmosphere adjustment at the stage of performing perfusion culture only in the central internal space S1 (first space S11), the gas atmosphere adjustment at the stage in which the peripheral internal space S2 was communicated was This can be done using the first channel 311 and the second channel 343 .
　また、上記のように、複数のポート１９を介して周囲内部空間Ｓ２に対する培養液Ｃの注入および排出が可能であれば、図８に示すように、吸着パッド５２の第１底面４１ａへの密着状態を維持したまま、あらかじめポート１９を介して周囲内部空間Ｓ２に適当量の培養液Ｃを供給することができる。この場合、その後に吸着パッド５２の密着状態を解放すると、培養液Ｃが周囲内部空間Ｓ２に急激に流入することは防止され、培養液Ｃの急激な流れによって生じる乱流を抑制することができる。このことは、拡大培養への適切な移行に寄与する。 Further, if the culture solution C can be injected into and discharged from the surrounding internal space S2 through the plurality of ports 19 as described above, the adhesion of the suction pad 52 to the first bottom surface 41a as shown in FIG. An appropriate amount of culture medium C can be supplied to the surrounding internal space S2 through the port 19 in advance while maintaining the state. In this case, when the suction pad 52 is released from the adhered state after that, the culture medium C is prevented from suddenly flowing into the surrounding internal space S2, and turbulence caused by the rapid flow of the culture medium C can be suppressed. . This contributes to proper transfer to expansion culture.
　吸着パッド５２の密着状態を解除する際、例えば吸引ポート５３（吸引路５３２）に対してポンプ（図示略）などによって短時間送気することで、吸着パッド５２の吸着状態を迅速、且つ容易に解除することができる。これにより、極めてシンプルな機構で培養容器の容積拡大の自動化が実現できる。 When releasing the close contact state of the suction pad 52, for example, by supplying air to the suction port 53 (suction path 532) for a short period of time using a pump (not shown), the suction state of the suction pad 52 can be quickly and easily removed. can be released. This makes it possible to automate the expansion of the volume of the culture vessel with an extremely simple mechanism.
　＜第２実施形態＞
　図１０～図１２は、本開示の第２実施形態に係る細胞培養用容器を示している。なお、図１０以降の図面において、上記実施形態の細胞培養用容器Ａ１と同一または類似の要素には、上記実施形態と同一の符号を付しており、適宜説明を省略する。 <Second embodiment>
10 to 12 show a cell culture vessel according to a second embodiment of the present disclosure. In addition, in the drawings after FIG. 10 , the same or similar elements as those of the cell culture vessel A1 of the above embodiment are denoted by the same reference numerals as those of the above embodiment, and description thereof will be omitted as appropriate.
　本実施形態の細胞培養用容器Ａ２は、上記実施形態の細胞培養用容器Ａ１と比べて、主に吸着部材５の構成が異なっている。 The cell culture vessel A2 of this embodiment differs from the cell culture vessel A1 of the above embodiment mainly in the structure of the adsorption member 5.
　本実施形態において、吸着部材５は柱状部５４をさらに有する。柱状部５４は、吸引ポート５３から離れて配置されており、図示した例では、吸着部材５の周方向において吸引ポート５３から最も離れた位置に設けられている。柱状部５４は、ブロック部５１から第１方向一方側ｘ１に延びる円柱状とされており、横断面円形の外周面を有する。柱状部５４は、第１環状板部３１に形成された貫通孔３１４を通じて当該第１環状板部３１よりも第１方向一方側ｘ１に突出している。本実施形態において、図１２に示すように、柱状部５４の先端（第１方向一方側ｘ１端）には、柱状部５４の外周面よりも外向きに突出する突起部５４１が設けられている。貫通孔３１４の内径寸法は、自然状態において柱状部５４の外径寸法よりも僅かに小さくされている。柱状部５４は、貫通孔３１４に圧入されている。これにより、貫通孔３１４（第１環状板部３１）は、柱状部５４の外周面に密着（当接）して当該外周面を囲っている。 In this embodiment, the adsorption member 5 further has a columnar portion 54 . The columnar portion 54 is arranged away from the suction port 53 , and in the illustrated example, is provided at the farthest position from the suction port 53 in the circumferential direction of the adsorption member 5 . The columnar portion 54 has a columnar shape extending from the block portion 51 in the first direction one side x1 and has an outer peripheral surface with a circular cross section. The columnar portion 54 protrudes from the first annular plate portion 31 to the first direction one side x1 through a through hole 314 formed in the first annular plate portion 31 . In the present embodiment, as shown in FIG. 12 , a protrusion 541 that protrudes outward from the outer peripheral surface of the columnar portion 54 is provided at the tip of the columnar portion 54 (first direction one side x1 end). . The inner diameter of the through hole 314 is slightly smaller than the outer diameter of the columnar portion 54 in the natural state. The columnar portion 54 is press-fitted into the through hole 314 . As a result, the through hole 314 (the first annular plate portion 31) is in close contact with (contacts with) the outer peripheral surface of the columnar portion 54 to surround the outer peripheral surface.
　また、本実施形態では、図１２等から理解されるように、上下方向（第１方向）に見て、第１環状板部３１における貫通孔３１４の周囲は、柱状部５４の先端に設けられた突起部５４１と重なっている。突起部５４１は、第１環状板部３１の上方に隣接している。これにより、第１環状板部３１の上端（第１方向一方側ｘ１端）には、突起部５４１が当接し得る。 Moreover, in the present embodiment, as can be understood from FIG. It overlaps with the projecting portion 541 . The projecting portion 541 is adjacent above the first annular plate portion 31 . Thereby, the protrusion 541 can come into contact with the upper end (first direction one side x1 end) of the first annular plate portion 31 .
　本実施形態の細胞培養用容器Ａ２において、第１フィルム状部材１および第２フィルム状部材２は、各々が環状で外周部１２，２２どうしが互いにつながり、内周側の第１開口１１および第２開口２１が第１プレート体３および第２プレート体４によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ２によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture container A2 of the present embodiment, the first film-like member 1 and the second film-like member 2 are each annular, the outer peripheral portions 12 and 22 are connected to each other, and the first opening 11 on the inner peripheral side and the second film-like member 2 are connected to each other. Two openings 21 are blocked by the first plate body 3 and the second plate body 4 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A2 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　また、本実施形態では、吸着部材５は柱状部５４を有し、柱状部５４の先端（第１方向一方側ｘ１端）には、柱状部５４の外周面よりも外向きに突出する突起部５４１が設けられている。突起部５４１は、第１環状板部３１の上方に隣接しており、第１環状板部３１の上端に当接し得る。このような構成によれば、例えば拡大培養に移行する際に吸着パッド５２の吸着状態の解除が迅速に行われないと、第１プレート体３に対して吸着部材５が第１底面４１ａ（第２プレート体４）側である下方（第１方向他方側ｘ２）に引っ張られる力が作用する。このように吸着部材５に下方への力が作用しても、突起部５４１が第１環状板部３１の上端により係止されることで、吸着部材５が第１プレート体３から離脱することは防止される。また、吸着部材５の延出部５３１においても、延出部５３１の先端（第１方向一方側ｘ１端）には、延出部５３１の外周面よりも外向きに突出する突起部５３３が設けられている。これにより、吸着部材５に下方への力が作用しても、延出部５３１が第１環状板部３１の上端により係止されることで、吸着部材５が第１プレート体３から離脱することは防止される。なお、本実施形態では、吸着部材５は１つの柱状部５４を有する構成であるが、複数の柱状部５４を有する構成としてもよい。吸着部材５が複数の柱状部５４を有する場合、延出部５３１および複数の柱状部５４は、吸着部材５の周方向において均等に分散して配置するのが好ましい。 In addition, in the present embodiment, the adsorption member 5 has a columnar portion 54, and at the tip of the columnar portion 54 (first direction one side x1 end), there is a protrusion projecting outward from the outer peripheral surface of the columnar portion 54. 541 is provided. The projecting portion 541 is adjacent to the upper side of the first annular plate portion 31 and can abut on the upper end of the first annular plate portion 31 . According to such a configuration, if the adsorption state of the adsorption pad 52 is not quickly released when shifting to the expansion culture, for example, the adsorption member 5 will not move to the first bottom surface 41a (the first bottom surface 41a) of the first plate body 3 A pulling force acts downward (the second side in the first direction x2) on the 2 plate body 4) side. Even if a downward force acts on the attracting member 5 in this manner, the projecting portion 541 is locked by the upper end of the first annular plate portion 31 , so that the attracting member 5 is separated from the first plate body 3 . is prevented. Also, in the extension portion 531 of the adsorption member 5, a projection portion 533 projecting outward from the outer peripheral surface of the extension portion 531 is provided at the tip of the extension portion 531 (first direction one side x1 end). It is As a result, even if a downward force acts on the attracting member 5 , the extending portion 531 is locked by the upper end of the first annular plate portion 31 , so that the attracting member 5 is separated from the first plate body 3 . is prevented. Although the adsorption member 5 has one columnar portion 54 in this embodiment, it may have a plurality of columnar portions 54 . When the adsorption member 5 has a plurality of columnar portions 54 , it is preferable that the extension portion 531 and the plurality of columnar portions 54 are evenly distributed in the circumferential direction of the adsorption member 5 .
　その他にも、本実施形態の細胞培養用容器Ａ２において、上記実施形態の細胞培養用容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A2 of the present embodiment has the same effects as the cell culture vessel A1 of the above embodiment.
　＜第３実施形態＞
　図１３および図１４は、本開示の第３実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ３は、上記実施形態の細胞培養用容器Ａ１と比べて、第２プレート体４の構成が異なっている。 <Third Embodiment>
13 and 14 show a cell culture vessel according to a third embodiment of the present disclosure. The cell culture vessel A3 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second plate body 4 .
　本実施形態の細胞培養用容器Ａ３において、第２プレート体４（主板部４１）の第１底面４１ａには、複数の凹部４１１が形成されている。複数の凹部４１１は、第１底面４１ａにおいて、第１方向ｘに見て吸着パッド５２の内側の領域を含んで形成される。図１４に示すように、複数の凹部４１１は、第１底面４１ａの面内方向に沿って稠密に配列されている。複数の凹部４１１は、培養細胞が三次元的に凝集し、スフェロイドが形成されやすくするための凹みである。 In the cell culture vessel A3 of the present embodiment, a plurality of recesses 411 are formed in the first bottom surface 41a of the second plate body 4 (main plate portion 41). The plurality of recesses 411 are formed on the first bottom surface 41a so as to include regions inside the suction pads 52 when viewed in the first direction x. As shown in FIG. 14, the plurality of recesses 411 are densely arranged along the in-plane direction of the first bottom surface 41a. The plurality of recesses 411 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
　本実施形態の細胞培養用容器Ａ３において、第１フィルム状部材１および第２フィルム状部材２は、各々が環状で外周部１２，２２どうしが互いにつながり、内周側の第１開口１１および第２開口２１が第１プレート体３および第２プレート体４によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ３によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture container A3 of the present embodiment, the first film-like member 1 and the second film-like member 2 are each annular, the outer peripheral portions 12 and 22 are connected to each other, and the first opening 11 on the inner peripheral side and the second film-like member 2 are connected to each other. Two openings 21 are blocked by the first plate body 3 and the second plate body 4 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A3 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態では、主板部４１（第２プレート体４）の第１底面４１ａには複数の凹部４１１が設けられている。複数の凹部４１１を含む容器底部（第１底面４１ａ）には、スフェロイド形成に有利に働く細胞非接着コートを施してもよい。このような構成にすれば、細胞が三次元的に凝集し、スフェロイドが形成されやすい。本開示によれば、灌流の液中に気泡が残存しにくく、灌流スピードは極めてゆっくりしたものなのでスフェロイドが凹部４１１から不当に浮き上がりにくく、また、天井凹部３４２側に設けられた膜フィルター７によってスフェロイドが排出されることもない。上記のように中央内部空間Ｓ１（第１空間Ｓ１１）で灌流培養した細胞（スフェロイド）は、細胞培養用容器Ａ３から取り出すことなく、周囲内部空間Ｓ２を連通させた拡大培養へスムーズに移行することが可能となる。 In this embodiment, a plurality of recesses 411 are provided on the first bottom surface 41a of the main plate portion 41 (second plate body 4). A container bottom (first bottom surface 41a) including a plurality of recesses 411 may be coated with a non-adherent cell coating that favors spheroid formation. With such a configuration, cells are three-dimensionally aggregated, and spheroids are likely to be formed. According to the present disclosure, air bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the concave portion 411, and the membrane filter 7 provided on the ceiling concave portion 342 side prevents the spheroids from is not discharged. The cells (spheroids) perfused and cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture with the peripheral internal space S2 communicated without being removed from the cell culture container A3. becomes possible.
　その他にも、本実施形態の細胞培養用容器Ａ３において、上記実施形態の細胞培養用容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A3 of the present embodiment has the same effects as the cell culture vessel A1 of the above embodiment.
　＜第４実施形態＞
　図１５は、本開示の第４実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ４は、上記実施形態の細胞培養用容器Ａ１と比べて、主に第２バッグ部Ｂ２の構成が異なっている。 <Fourth Embodiment>
FIG. 15 shows a cell culture container according to a fourth embodiment of the present disclosure. The cell culture vessel A4 of the present embodiment differs from the cell culture vessel A1 of the above embodiment mainly in the configuration of the second bag portion B2.
　本実施形態の細胞培養用容器Ａ４において、上記実施形態と異なり、第２フィルム状部材２は、第２開口２１を有しておらず、略円形状である。一方、第２プレート体４は、略円環状とされている。第２プレート体４は、円環状の主板部４１を備える。主板部４１は、第１底面４１ａを有する。第１底面４１ａは、第１方向一方側ｘ１を向き、略平坦である。この第１底面４１ａは、吸着パッド５２が密着する部位である。第２プレート体４（主板部４１）の下面（第１方向他方側ｘ２を向く面）は、第２フィルム状部材２の中央部における第１方向一方側ｘ１を向く面と接合されている。 In the cell culture vessel A4 of this embodiment, unlike the above embodiment, the second film-like member 2 does not have the second opening 21 and has a substantially circular shape. On the other hand, the second plate body 4 has a substantially annular shape. The second plate body 4 includes an annular main plate portion 41 . The main plate portion 41 has a first bottom surface 41a. The first bottom surface 41a faces the first direction one side x1 and is substantially flat. The first bottom surface 41a is a portion to which the suction pad 52 is brought into close contact. The lower surface (the surface facing the other side x2 in the first direction) of the second plate body 4 (the main plate portion 41) is joined to the surface facing the one side x1 in the first direction in the central portion of the second film member 2.
　本実施形態の細胞培養用容器Ａ４において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ４によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。その他にも、本実施形態の細胞培養用容器Ａ４は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In the cell culture vessel A4 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A4 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume. In addition, the cell culture vessel A4 of the present embodiment has the same effects as those of the above embodiment within the same configuration range as the cell culture vessel A1 of the above embodiment.
　＜第５実施形態＞
　図１６は、本開示の第５実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ５は、上記実施形態の細胞培養用容器Ａ４と比べて、第２バッグ部Ｂ２の構成が異なっている。 <Fifth Embodiment>
FIG. 16 shows a cell culture container according to a fifth embodiment of the present disclosure. The cell culture vessel A5 of this embodiment differs from the cell culture vessel A4 of the above embodiment in the configuration of the second bag portion B2.
　本実施形態の細胞培養用容器Ａ５において、上記の細胞培養用容器Ａ４と異なり、第２プレート体４を具備していない。吸着パッド５２は、第２フィルム状部材２において第１方向一方側ｘ１を向く底面２３ａに密着可能である。 In the cell culture vessel A5 of this embodiment, unlike the cell culture vessel A4 described above, the second plate body 4 is not provided. The suction pad 52 can be brought into close contact with the bottom surface 23a of the second film member 2 facing the one side x1 in the first direction.
　本実施形態の細胞培養用容器Ａ４において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２フィルム状部材２の底面２３ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の底面２３ａに対する密着状態を解放し、吸着パッド５２が底面２３ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ４によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。その他にも、本実施形態の細胞培養用容器Ａ５は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In the cell culture vessel A4 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 with respect to the bottom surface 23a so that the suction pad 52 does not adhere to the bottom surface 23a, the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate. According to the cell culture vessel A4 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume. In addition, the cell culture vessel A5 of the present embodiment has the same effects as those of the above embodiment within the same range of configuration as the cell culture vessel A1 of the above embodiment.
　＜第６実施形態＞
　図１７は、本開示の第６実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ６は、上記実施形態の細胞培養用容器Ａ４と比べて、第２フィルム状部材２の構成が異なっている。 <Sixth embodiment>
FIG. 17 shows a cell culture container according to a sixth embodiment of the present disclosure. The cell culture vessel A6 of the present embodiment differs from the cell culture vessel A4 of the above embodiment in the configuration of the second film-like member 2 .
　本実施形態の細胞培養用容器Ａ６においては、上記の細胞培養用容器Ａ４と比べて、第２フィルム状部材２の構成が異なっている。第２フィルム状部材２において第１方向一方側ｘ１を向く底面２３ａには、複数の凹部２３１が形成されている。複数の凹部２３１は、底面２３ａのうち第１方向ｘに見て吸着パッド５２の内側の領域を含んで形成される。詳細な図示は省略するが、複数の凹部２３１は、図１４に示した複数の凹部４１１と同様に、底面２３ａの面内方向に沿って稠密に配列されている。複数の凹部２３１は、培養細胞が三次元的に凝集し、スフェロイドが形成されやすくするための凹みである。 In the cell culture vessel A6 of this embodiment, the configuration of the second film member 2 is different from that of the cell culture vessel A4. A plurality of recesses 231 are formed in the bottom surface 23a of the second film member 2 facing the first direction one side x1. The plurality of recesses 231 are formed including areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 23a. Although detailed illustration is omitted, the plurality of recesses 231 are densely arranged along the in-plane direction of the bottom surface 23a in the same manner as the plurality of recesses 411 shown in FIG. The plurality of recesses 231 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
　本実施形態の細胞培養用容器Ａ６において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ６によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A6 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A6 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) is shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態では、第２フィルム状部材２の底面２３ａには複数の凹部２３１が設けられている。複数の凹部２３１を含む容器底部（底面２３ａ）には、スフェロイド形成に有利に働く細胞非接着コートを施してもよい。このような構成にすれば、細胞が三次元的に凝集し、スフェロイドが形成されやすい。本開示によれば、灌流の液中に気泡が残存しにくく、灌流スピードは極めてゆっくりしたものなのでスフェロイドが凹部２３１から不当に浮き上がりにくく、また、天井凹部３４２側に設けられた膜フィルター７によってスフェロイドが排出されることもない。上記のように中央内部空間Ｓ１（第１空間Ｓ１１）で灌流培養した細胞（スフェロイド）は、細胞培養用容器Ａ６から取り出すことなく、周囲内部空間Ｓ２を連通させた拡大培養へスムーズに移行することが可能となる。その他にも、本実施形態の細胞培養用容器Ａ６は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In this embodiment, the bottom surface 23a of the second film member 2 is provided with a plurality of recesses 231. A container bottom (bottom surface 23a) including a plurality of recesses 231 may be coated with a non-adhesive cell coating that favors spheroid formation. With such a configuration, cells are three-dimensionally aggregated, and spheroids are likely to be formed. According to the present disclosure, bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the recess 231, and the membrane filter 7 provided on the ceiling recess 342 side prevents spheroids from is not discharged. The cells (spheroids) that have been perfusion cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture in which the surrounding internal space S2 is communicated without being removed from the cell culture container A6. becomes possible. In addition, the cell culture vessel A6 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
　＜第７実施形態＞
　図１８は、本開示の第７実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ７は、上記実施形態の細胞培養用容器Ａ５と比べて、第２フィルム状部材２の構成が異なっている。 <Seventh Embodiment>
FIG. 18 shows a cell culture container according to a seventh embodiment of the present disclosure. The cell culture vessel A7 of this embodiment differs from the cell culture vessel A5 of the above embodiment in the configuration of the second film member 2 .
　本実施形態において、第２フィルム状部材２において第１方向一方側ｘ１を向く底面２３ａには、複数の凹部２３１が形成されている。複数の凹部２３１は、底面２３ａのうち第１方向ｘに見て吸着パッド５２の内側の領域を含んで形成される。詳細な図示は省略するが、複数の凹部２３１は、図１４に示した複数の凹部４１１と同様に、底面２３ａの面内方向に沿って稠密に配列されている。複数の凹部２３１は、培養細胞が三次元的に凝集し、スフェロイドが形成されやすくするための凹みである。 In the present embodiment, a plurality of recesses 231 are formed on the bottom surface 23a of the second film-like member 2 facing the one side x1 in the first direction. The plurality of recesses 231 are formed including areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 23a. Although detailed illustration is omitted, the plurality of recesses 231 are densely arranged along the in-plane direction of the bottom surface 23a in the same manner as the plurality of recesses 411 shown in FIG. The plurality of recesses 231 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
　本実施形態の細胞培養用容器Ａ７において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２フィルム状部材２の底面２３ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の底面２３ａに対する密着状態を解放し、吸着パッド５２が底面２３ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ７によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A7 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 with respect to the bottom surface 23a so that the suction pad 52 does not adhere to the bottom surface 23a, the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate. According to the cell culture vessel A7 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態では、第２フィルム状部材２の底面２３ａには複数の凹部２３１が設けられている。複数の凹部２３１を含む容器底部（底面２３ａ）には、スフェロイド形成に有利に働く細胞非接着コートを施してもよい。このような構成にすれば、細胞が三次元的に凝集し、スフェロイドが形成されやすい。本開示によれば、灌流の液中に気泡が残存しにくく、灌流スピードは極めてゆっくりしたものなのでスフェロイドが凹部２３１から不当に浮き上がりにくく、また、天井凹部３４２側に設けられた膜フィルター７によってスフェロイドが排出されることもない。上記のように中央内部空間Ｓ１（第１空間Ｓ１１）で灌流培養した細胞（スフェロイド）は、細胞培養用容器Ａ７から取り出すことなく、周囲内部空間Ｓ２を連通させた拡大培養へスムーズに移行することが可能となる。その他にも、本実施形態の細胞培養用容器Ａ７は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In this embodiment, the bottom surface 23a of the second film member 2 is provided with a plurality of recesses 231. A container bottom (bottom surface 23a) including a plurality of recesses 231 may be coated with a non-adhesive cell coating that favors spheroid formation. With such a configuration, cells are three-dimensionally aggregated, and spheroids are likely to be formed. According to the present disclosure, bubbles are less likely to remain in the perfusion liquid, and the perfusion speed is extremely slow, so the spheroids are less likely to unduly rise from the recess 231, and the membrane filter 7 provided on the ceiling recess 342 side prevents spheroids from is not discharged. The cells (spheroids) that have been perfusion cultured in the central internal space S1 (first space S11) as described above can be smoothly transferred to expansion culture with the surrounding internal space S2 communicated without being removed from the cell culture container A7. becomes possible. In addition, the cell culture vessel A7 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
　＜第８実施形態＞
　図１９は、本開示の第８実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ８は、上記実施形態の細胞培養用容器Ａ１と比べて、第２バッグ部Ｂ２の構成が異なっている。 <Eighth Embodiment>
FIG. 19 shows a cell culture container according to an eighth embodiment of the present disclosure. The cell culture vessel A8 of the present embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second bag portion B2.
　本実施形態の第２バッグ部Ｂ２において、第２プレート体４は、第２フィルム状部材２に取り付けられておらず、第２フィルム状部材とは別体となっている。また、第２プレート体４の面積は大きくされており、第１方向ｘに見て、第２プレート体４の面積は、第１プレート体３よりも大きい。 In the second bag portion B2 of this embodiment, the second plate body 4 is not attached to the second film-like member 2 and is separate from the second film-like member. Further, the area of the second plate body 4 is made large, and the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x.
　本実施形態の細胞培養用容器Ａ８において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ８によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A8 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A8 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態では、第２バッグ部Ｂ２は、第２フィルム状部材２とは別体とされた第２プレート体４を含む。接着細胞の培養に適した素材として例えばポリカーボネートが挙げられるが、第２プレート体４の構成材料がポリカーボネートである場合、第２プレート体４と第２フィルム状部材２との接着性が低下するおそれがある。本実施形態の構成によれば、第２プレート体４を第２フィルム状部材２とは別体とすることで、第２プレート体４の素材を目的に応じて適宜選択することができる。また、本実施形態において、第１方向ｘに見て、第２プレート体４の面積は、第１プレート体３よりも大きい。このような構成によれば、拡大培養に移行した後においても、大面積の第２プレート体４上において細胞培養を適切に継続することができる。その他にも、本実施形態の細胞培養用容器Ａ８は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In this embodiment, the second bag portion B2 includes a second plate body 4 that is separate from the second film-like member 2 . Materials suitable for culturing adherent cells include, for example, polycarbonate. However, if the material constituting the second plate member 4 is polycarbonate, there is a possibility that the adhesiveness between the second plate member 4 and the second film-like member 2 may be reduced. There is According to the configuration of the present embodiment, by making the second plate body 4 separate from the second film-like member 2, the material of the second plate body 4 can be appropriately selected according to the purpose. Moreover, in the present embodiment, the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x. According to such a configuration, cell culture can be appropriately continued on the large-area second plate body 4 even after shifting to expansion culture. In addition, the cell culture vessel A8 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
　＜第９実施形態＞
　図２０は、本開示の第９実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ９は、上記実施形態の細胞培養用容器Ａ１と比べて、第２プレート体４の構成が異なっている。 <Ninth Embodiment>
FIG. 20 shows a cell culture container according to the ninth embodiment of the present disclosure. The cell culture vessel A9 of the present embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second plate body 4 .
　本実施形態において、第２プレート体４の面積が大きくされている。第１方向ｘに見て、第２プレート体４の面積は、第１プレート体３よりも大きい。 In this embodiment, the area of the second plate body 4 is increased. The area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x.
　本実施形態の細胞培養用容器Ａ９において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ９によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A9 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A9 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　また、本実施形態において、第１方向ｘに見て、第２プレート体４の面積は、第１プレート体３よりも大きい。このような構成によれば、拡大培養に移行した後においても、大面積の第２プレート体４上において細胞培養を適切に継続することができる。その他にも、本実施形態の細胞培養用容器Ａ９は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 Also, in the present embodiment, the area of the second plate body 4 is larger than that of the first plate body 3 when viewed in the first direction x. According to such a configuration, cell culture can be appropriately continued on the large-area second plate body 4 even after shifting to expansion culture. In addition, the cell culture vessel A9 of the present embodiment has the same effect as the above embodiment within the same range of configuration as the cell culture vessel A1 of the above embodiment.
　＜第１０実施形態＞
　図２１は、本開示の第１０実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ１０は、上記実施形態の細胞培養用容器Ａ１と比べて、第３フィルム部材９をさらに備え、これに伴い適宜変更が加えられている。 <Tenth Embodiment>
FIG. 21 shows a cell culture container according to a tenth embodiment of the present disclosure. Compared to the cell culture vessel A1 of the above embodiment, the cell culture vessel A10 of the present embodiment further includes a third film member 9, and is appropriately modified accordingly.
　細胞培養用容器Ａ１０において、第３フィルム部材９は、第２フィルム状部材２（第２バッグ部Ｂ２）に対して第１方向他方側ｘ２に位置する。第３フィルム部材９の外周部９２は、第１フィルム状部材１の外周部１２および第２フィルム状部材２の外周部２２とともに例えば熱溶着等の適宜手段によって接合され、互いにつながっている。第２バッグ部Ｂ２（第２フィルム状部材２および第２プレート体４）の第１方向他方側ｘ２（図２１の下方）と、第３フィルム部材９との間には、第４空間Ｓ４が形成されている。第２フィルム状部材２において、外周部１２寄りの適所には、１または複数の連通口２４が形成されている。連通口２４は、第２フィルム状部材２の厚さ方向に貫通しており、この連通口２４を介して周囲内部空間Ｓ２と第４空間Ｓ４とが連通している。また、吸着パッド５２が第２プレート体４の第１底面４１ａに密着しない状態にあるとき、中央内部空間Ｓ１と周囲内部空間Ｓ２とが連通する。これにより、吸着パッド５２が第１底面４１ａに密着しない状態にあるとき、中央内部空間Ｓ１と第４空間Ｓ４とが連通口２４を介して連通する。上記の連通口２４は、「連通手段」の一例に相当する。 In the cell culture container A10, the third film member 9 is positioned on the other side x2 in the first direction with respect to the second film member 2 (second bag portion B2). The outer peripheral portion 92 of the third film member 9 is joined together with the outer peripheral portion 12 of the first film member 1 and the outer peripheral portion 22 of the second film member 2 by an appropriate means such as heat welding, and connected to each other. A fourth space S4 is provided between the second bag portion B2 (the second film member 2 and the second plate member 4) on the other side x2 in the first direction (lower side in FIG. 21) and the third film member 9. formed. In the second film-like member 2, one or a plurality of communication holes 24 are formed at appropriate locations near the outer peripheral portion 12. As shown in FIG. The communication port 24 penetrates through the second film member 2 in the thickness direction, and the surrounding internal space S2 and the fourth space S4 are communicated through the communication port 24 . Also, when the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate body 4, the central internal space S1 and the peripheral internal space S2 communicate with each other. Thereby, when the suction pad 52 is not in close contact with the first bottom surface 41a, the central internal space S1 and the fourth space S4 communicate with each other through the communication port 24. As shown in FIG. The communication port 24 described above corresponds to an example of "communication means".
　本実施形態では、第３フィルム部材９において第１方向一方側ｘ１を向く底面９３ａには、複数の凹部９３１が形成されている。複数の凹部９３１は、底面９３ａのうち第１方向ｘに見て吸着パッド５２の内側の領域を含んで形成される。詳細な図示は省略するが、複数の凹部９３１は、図１４に示した複数の凹部４１１と同様に、底面９３ａの面内方向に沿って稠密に配列されている。複数の凹部９３１は、培養細胞が三次元的に凝集し、スフェロイドが形成されやすくするための凹みである。 In the present embodiment, a plurality of recesses 931 are formed on the bottom surface 93a of the third film member 9 facing the one side x1 in the first direction. The plurality of recesses 931 are formed to include areas inside the suction pads 52 when viewed in the first direction x on the bottom surface 93a. Although detailed illustration is omitted, the plurality of recesses 931 are densely arranged along the in-plane direction of the bottom surface 93a, like the plurality of recesses 411 shown in FIG. The plurality of recesses 931 are recesses for facilitating the three-dimensional aggregation of cultured cells and the formation of spheroids.
　本実施形態の細胞培養用容器Ａ１０において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ１０によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A10 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A10 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態の細胞培養用容器Ａ１０は、第３フィルム部材９を備え、第２バッグ部Ｂ２の第１方向他方側ｘ２と第３フィルム部材９とに挟まれた第４空間Ｓ４を有する。吸着パッド５２が第２プレート体４の第１底面４１ａ（第２バッグ部Ｂ２）に密着しない状態にあるとき、中央内部空間Ｓ１と第４空間Ｓ４とが連通口２４を介して連通する。このような構成によれば、たとえば吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態（拡大培養養前の前培養）では、第２プレート体４の第１底面４１ａを利用して接着細胞の培養を行う。吸着パッド５２が第２プレート体４の第１底面４１ａに密着しない状態（拡大培養）では、培養細胞が連通口２４を介して第４空間Ｓ４に移動し、第３フィルム部材９の底面９３ａに形成された凹部９３１に収容される。ここでは、培養細胞が三次元的に凝集し、スフェロイドが形成されやすい。このように、本実施形態によれば、前培養と拡大培養とで異なった性質の異なる培養を行うことが可能である。その他にも、本実施形態の細胞培養用容器Ａ１０は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 The cell culture vessel A10 of the present embodiment includes a third film member 9 and has a fourth space S4 sandwiched between the third film member 9 and the first direction other side x2 of the second bag portion B2. When the suction pad 52 is not in close contact with the first bottom surface 41a (the second bag portion B2) of the second plate body 4, the central internal space S1 and the fourth space S4 communicate with each other through the communication port 24. According to such a configuration, for example, when the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate body 4 (preculture before expansion culture), the first Adherent cells are cultured using the bottom surface 41a. When the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate member 4 (expansion culture), the cultured cells move to the fourth space S4 through the communication port 24 and reach the bottom surface 93a of the third film member 9. It is accommodated in the formed recess 931 . Here, cultured cells tend to aggregate three-dimensionally and form spheroids. Thus, according to the present embodiment, it is possible to perform culture with different properties between the preculture and the expansion culture. In addition, the cell culture vessel A10 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
　＜第１１実施形態＞
　図２２は、本開示の第１１実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ１１は、上記実施形態の細胞培養用容器Ａ１０と比べて、第２フィルム状部材２に設けられた連通口２４に代えて第３フィルム部材９にポート９９が設けられており、これに伴い適宜変更が加えられている。 <Eleventh Embodiment>
FIG. 22 shows a cell culture container according to the eleventh embodiment of the present disclosure. Unlike the cell culture vessel A10 of the above embodiment, the cell culture vessel A11 of the present embodiment has a port 99 provided on the third film member 9 instead of the communication port 24 provided on the second film member 2. It has been modified accordingly.
　細胞培養用容器Ａ１１において、第３フィルム部材９には、複数のポート９９が設けられている。第３フィルム部材９において各ポート９９に対応する箇所には、ポート用孔９４が形成されている。各ポート１９は、一方側端（第１方向他方側ｘ２端）が外部に通じており、他方側端（第１方向一方側ｘ１端）が第３フィルム部材９のポート９９を介して第４空間Ｓ４２に通じている。本実施形態では、第１フィルム状部材１側のポート１９および第３フィルム部材９のポート９９には、チューブ８８が接続されている。チューブ８８の一方側端は、ポート１９に接続されており、チューブ８８の他方側端はポート９９に接続されている。第２フィルム状部材２においては、細胞培養用容器Ａ１０と異なり、連通口２４が形成されていない。なお、図２２においては、チューブ８８の中間部分を省略している。 In the cell culture container A11, the third film member 9 is provided with a plurality of ports 99. Port holes 94 are formed at portions of the third film member 9 corresponding to the respective ports 99 . Each port 19 has one side end (first direction other side x2 end) communicating with the outside, and the other side end (first direction one side x1 end) communicates with the fourth through port 99 of third film member 9 . It leads to space S42. In this embodiment, a tube 88 is connected to the port 19 of the first film member 1 and the port 99 of the third film member 9 . One end of tube 88 is connected to port 19 and the other end of tube 88 is connected to port 99 . In the second film member 2, unlike the cell culture vessel A10, the communication port 24 is not formed. 22, an intermediate portion of the tube 88 is omitted.
　上記構成により、チューブ８８を介して周囲内部空間Ｓ２と第４空間Ｓ４とが連通している。また、吸着パッド５２が第２プレート体４の第１底面４１ａに密着しない状態にあるとき、中央内部空間Ｓ１と周囲内部空間Ｓ２とが連通する。これにより、吸着パッド５２が第１底面４１ａに密着しない状態にあるとき、中央内部空間Ｓ１と第４空間Ｓ４とがポート１９、チューブ８８およびポート９９を介して連通する。上記のポート１９、チューブ８８およびポート９９は、「連通手段」の一例に相当する。 With the above configuration, the surrounding internal space S2 and the fourth space S4 are communicated via the tube 88. Also, when the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate body 4, the central internal space S1 and the peripheral internal space S2 communicate with each other. Thereby, the central internal space S1 and the fourth space S4 communicate with each other through the port 19, the tube 88 and the port 99 when the suction pad 52 is not in close contact with the first bottom surface 41a. The port 19, tube 88 and port 99 described above correspond to an example of "communication means".
　本実施形態の細胞培養用容器Ａ１１において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、吸着パッド５２が第１底面４１ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ１１によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A11 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate member 4, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is , the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 from the first bottom surface 41a and making the suction pad 52 not in close contact with the first bottom surface 41a, the central inner space S1 (first space S11) and the surrounding inner space are separated. It communicates with the space S2. According to the cell culture vessel A11 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, it is possible to shift to expansion culture during cell culture (perfusion culture) without changing the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　本実施形態の細胞培養用容器Ａ１１は、第３フィルム部材９を備え、第２バッグ部Ｂ２の第１方向他方側ｘ２と第３フィルム部材９とに挟まれた第４空間Ｓ４を有する。吸着パッド５２が第２プレート体４の第１底面４１ａ（第２バッグ部Ｂ２）に密着しない状態にあるとき、中央内部空間Ｓ１と第４空間Ｓ４とがポート１９、チューブ８８およびポート９９を介して連通する。このような構成によれば、たとえば吸着部材５の吸着パッド５２が第２プレート体４の第１底面４１ａに密着する状態（拡大培養養前の前培養）では、第２プレート体４の第１底面４１ａを利用して接着細胞の培養を行う。吸着パッド５２が第２プレート体４の第１底面４１ａに密着しない状態（拡大培養）では、培養細胞がポート１９、チューブ８８およびポート９９を介して第４空間Ｓ４に移動し、第３フィルム部材９の底面９３ａに形成された凹部９３１に収容される。ここでは、培養細胞が三次元的に凝集し、スフェロイドが形成されやすい。このように、本実施形態によれば、前培養と拡大培養とで異なった性質の異なる培養を行うことが可能である。その他にも、本実施形態の細胞培養用容器Ａ１１は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 The cell culture vessel A11 of the present embodiment includes a third film member 9 and has a fourth space S4 sandwiched between the third film member 9 and the first direction other side x2 of the second bag portion B2. When the suction pad 52 is not in close contact with the first bottom surface 41a (second bag portion B2) of the second plate member 4, the central internal space S1 and the fourth space S4 are connected through the port 19, the tube 88 and the port 99. communicate with. According to such a configuration, for example, when the suction pad 52 of the suction member 5 is in close contact with the first bottom surface 41a of the second plate body 4 (preculture before expansion culture), the first Adherent cells are cultured using the bottom surface 41a. When the suction pad 52 is not in close contact with the first bottom surface 41a of the second plate body 4 (expansion culture), the cultured cells move to the fourth space S4 through the port 19, the tube 88 and the port 99, and are transferred to the third film member. 9 is housed in a recess 931 formed in the bottom surface 93a of . Here, cultured cells tend to aggregate three-dimensionally and form spheroids. Thus, according to the present embodiment, it is possible to perform culture with different properties between the preculture and the expansion culture. In addition, the cell culture vessel A11 of the present embodiment has the same effect as the above embodiment within the same configuration as the cell culture vessel A1 of the above embodiment.
　＜第１２実施形態＞
　図２３は、本開示の第１２実施形態に係る細胞培養用容器を示している。本実施形態の細胞培養用容器Ａ１２は、上記実施形態の細胞培養用容器Ａ１と比べて、第２バッグ部Ｂ２の構成が異なっている。 <Twelfth Embodiment>
FIG. 23 shows a cell culture vessel according to a twelfth embodiment of the present disclosure. The cell culture vessel A12 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the second bag portion B2.
　本実施形態の第２バッグ部Ｂ２において、第２プレート体４は、第２フィルム状部材２に対して第１方向他方側ｘ２に位置し、略円形状である。第２フィルム状部材２は第２開口２１を有しておらず、略円形状である。第２プレート体４および第２フィルム状部材２は、第１方向ｘに見て中央内部空間Ｓ１と重なる。第２プレート体４の第１底面４１ａ（第１方向一方側ｘ１を向く面）には、例えばヒートシールや接着などの手法により、第２フィルム状部材２が接合されている。 In the second bag portion B2 of the present embodiment, the second plate body 4 is positioned on the other side x2 in the first direction with respect to the second film member 2 and has a substantially circular shape. The second film member 2 does not have the second opening 21 and has a substantially circular shape. The second plate member 4 and the second film member 2 overlap the central internal space S1 when viewed in the first direction x. The second film-like member 2 is joined to the first bottom surface 41a (the surface facing the one side x1 in the first direction) of the second plate member 4 by, for example, heat sealing or adhesion.
　本実施形態の細胞培養用容器Ａ１２において、第１フィルム状部材１および第２フィルム状部材２は、各々の外周部１２，２２どうしが互いにつながる。第１フィルム状部材１の内周側の第１開口１１は、第１プレート体３によって塞がれている。吸着部材５の吸着パッド５２が第２フィルム状部材２の底面２３ａに密着する状態では、第１フィルム状部材１、第２フィルム状部材２および吸着部材５で囲まれた周囲内部空間Ｓ２は、中央内部空間Ｓ１（第１空間Ｓ１１）と遮断されている。上記吸着部材５の吸着パッド５２の底面２３ａに対する密着状態を解放し、吸着パッド５２が底面２３ａに密着しない状態とすることで、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とが連通する。このような構成の細胞培養用容器Ａ１２によれば、上記実施形態の細胞培養用容器Ａ１と同様に、容器の移し替えを伴わずに、細胞培養（灌流培養）の途中で拡大培養に移行し、細胞培養（灌流培養）を継続することが可能である。したがって、容器を移し替える際に生じるコンタミネーションのリスクを無くし、容積を拡大した培養容器で行う拡大培養にスムーズに移行することができる。 In the cell culture vessel A12 of the present embodiment, the outer peripheral portions 12 and 22 of the first film-like member 1 and the second film-like member 2 are connected to each other. The first opening 11 on the inner peripheral side of the first film member 1 is closed by the first plate body 3 . When the suction pad 52 of the suction member 5 is in close contact with the bottom surface 23a of the second film member 2, the surrounding internal space S2 surrounded by the first film member 1, the second film member 2 and the suction member 5 is It is cut off from the central internal space S1 (first space S11). By releasing the contact state of the suction pad 52 of the suction member 5 with respect to the bottom surface 23a so that the suction pad 52 does not adhere to the bottom surface 23a, the central internal space S1 (first space S11) and the surrounding internal space S2 are separated. communicate. According to the cell culture vessel A12 having such a configuration, similarly to the cell culture vessel A1 of the above-described embodiment, the cell culture (perfusion culture) can be shifted to the expansion culture in the middle of the cell culture (perfusion culture) without transferring the vessel. , it is possible to continue cell culture (perfusion culture). Therefore, it is possible to eliminate the risk of contamination that occurs when the container is transferred, and smoothly transition to expansion culture performed in a culture container with an enlarged volume.
　細胞培養用容器Ａ１２において、第２プレート体４は第２フィルム状部材２に取り付けられており、第２フィルム状部材２に対して第１方向他方側ｘ２に位置する。このような構成によれば、吸着パッド５２が接する第２フィルム状部材２の底面２３ａを、第２プレート体４の第１底面４１ａに対応する略平坦面に維持することができる。これにより、吸着パッド５２が底面２３ａに密着する時、当該底面２３ａへの吸着パッド５２の密着状態は適切に維持される。また、底面２３ａにおいて、第１方向ｘに見て中央内部空間Ｓ１と重なる領域についても、第２プレート体４の第１底面４１ａに対応する略平坦面に維持することができる。これにより、上記底面２３ａを細胞培養に適した平面状態に維持することができる。その他にも、本実施形態の細胞培養用容器Ａ１２は、上記実施形態の細胞培養用容器Ａ１と同様の構成の範囲において、上記実施形態と同様の作用効果を奏する。 In the cell culture vessel A12, the second plate body 4 is attached to the second film-like member 2 and positioned on the other side x2 in the first direction with respect to the second film-like member 2. With such a configuration, the bottom surface 23 a of the second film-like member 2 with which the suction pad 52 contacts can be maintained as a substantially flat surface corresponding to the first bottom surface 41 a of the second plate body 4 . As a result, when the suction pad 52 is brought into close contact with the bottom surface 23a, the close contact state of the suction pad 52 with the bottom surface 23a is appropriately maintained. In addition, the region of the bottom surface 23a that overlaps the central internal space S1 when viewed in the first direction x can also be maintained as a substantially flat surface corresponding to the first bottom surface 41a of the second plate body 4 . Thereby, the bottom surface 23a can be maintained in a planar state suitable for cell culture. In addition, the cell culture vessel A12 of the present embodiment has the same effect as the above embodiment within the same configuration range as the cell culture vessel A1 of the above embodiment.
　以上、本発明の具体的な実施形態を説明したが、本発明はこれに限定されるものではなく、発明の思想から逸脱しない範囲内で種々な変更が可能である。本発明に係る細胞培養用容器の各部の具体的な構成は、種々に設計変更自在である。 Although a specific embodiment of the present invention has been described above, the present invention is not limited to this, and various modifications are possible without departing from the spirit of the invention. The specific configuration of each part of the cell culture vessel according to the present invention can be changed in various ways.
　本開示の細胞培養用容器は、上記の各実施形態で示した略円形の構成に限定されず、例えば略長矩形でもよい。図２４は、略長矩形に構成された細胞培養用容器Ａ１３を示している。このような構成によれば、上記の第１空間Ｓ１１で培養した細胞を例えばシート状に形成した後、上記吸着部材５の吸着パッド５２の第１底面４１ａに対する密着状態を解放し、中央内部空間Ｓ１（第１空間Ｓ１１）と周囲内部空間Ｓ２とを連通させる。そうすると、図２５に示すように、シート状細胞Ｓｃを細胞培養用容器Ａ１３の一方端（図中右端）に寄せることが可能である。その後、図２６に示すように、シート状細胞Ｓｃが内包されるようにヒートシールにより第１フィルム状部材１および第２フィルム状部材２おいてヒートシール部Ｈｓを形成し、当該ヒートシール部Ｈｓを切り離す。これにより、形成されたシート状細胞Ｓｃを上記細胞培養用容器Ａ１３から取り出すことなく、閉鎖的にパッケージングできる。したがって、上記の細胞培養用容器Ａ１３は、作成したシート状細胞Ｓｃをコンタミネーションのリスクなく輸送、保管をするのに都合がよい。また、本開示の細胞培養用容器は、１つの第２バッグ部に複数の吸着パッドが密着し、各吸着パッドに対応する複数の中央内部空間が形成されるように構成してもよい。 The cell culture vessel of the present disclosure is not limited to the substantially circular configuration shown in each of the above embodiments, and may be, for example, substantially rectangular. FIG. 24 shows a cell culture vessel A13 configured in a substantially rectangular shape. According to such a configuration, after the cells cultured in the first space S11 are formed into, for example, a sheet shape, the suction member 5 is released from the adhesion state of the suction pad 52 to the first bottom surface 41a, and the central inner space is removed. S1 (first space S11) and surrounding internal space S2 are communicated. Then, as shown in FIG. 25, the sheet-like cells Sc can be moved to one end (right end in the figure) of the cell culture container A13. After that, as shown in FIG. 26, a heat-sealed portion Hs is formed in the first film-like member 1 and the second film-like member 2 by heat-sealing such that the sheet-like cells Sc are included, and the heat-sealed portion Hs is formed. detach the As a result, the formed sheet-like cells Sc can be closedly packaged without being removed from the cell culture container A13. Therefore, the cell culture container A13 is convenient for transporting and storing the prepared sheet-like cells Sc without risk of contamination. Further, the cell culture container of the present disclosure may be configured such that a plurality of suction pads are in close contact with one second bag portion, and a plurality of central internal spaces corresponding to each suction pad are formed.
　Ａ１，Ａ２，Ａ３，Ａ４，Ａ５，Ａ６，Ａ７，Ａ８，Ａ９，Ａ１０，Ａ１１，Ａ１２，Ａ１３：細胞培養用容器、Ｂ１：第１バッグ部、Ｂ２：第２バッグ部、１：第１フィルム状部材、１１：第１開口、１２：外周部、１３：ポート用孔、２：第２フィルム状部材、２１：第２開口、２２：外周部、２３ａ：底面、２３１：凹部、２４：連通口、３：第１プレート体、３１：第１環状板部、３１１：第１流路、３１２：吸着部材用凹溝、３１３，３１４：貫通孔、３２：第２環状板部、３３：膨出部、３３１：第３流路、３４：フィルター押え、３４１：筒状部、３４２：天井凹部、３４３：第２流路、４：第２プレート体、４１：主板部、４１ａ：第１底面、４１１：凹部、５：吸着部材、５１：ブロック部、５２：吸着パッド、５３：吸引ポート、５３１：延出部、５３２：吸引路、５３３：突起部、５４：柱状部、５４１：突起部、６：ガス透過膜、７：膜フィルター、８１，８２，８３：チューブ、８４：廃液排出口、８５：フィルター、８８：チューブ、９：第３フィルム部材、９２：外周部、９３ａ：底面、９３１：凹部、９４：ポート用孔、９９：ポート、Ｃ：培養液、Ｓ１：中央内部空間、Ｓ１１：第１空間、Ｓ１２：第２空間、Ｓ１３：第３空間、Ｓ２：周囲内部空間、Ｓ４：第４空間、Ｓｃ：シート状細胞、Ｈｓ：ヒートシール部、ｘ：第１方向、ｘ１：第１方向一方側、ｘ２：第１方向他方側 A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13: cell culture vessel, B1: first bag portion, B2: second bag portion, 1: first film 11: First opening 12: Peripheral portion 13: Port hole 2: Second film-like member 21: Second opening 22: Peripheral portion 23a: Bottom surface 231: Concave portion 24: Communication Mouth 3: first plate body 31: first annular plate portion 311: first flow path 312: concave groove for adsorption member 313, 314: through hole 32: second annular plate portion 33: swelling Outlet 331: Third flow path 34: Filter holder 341: Cylindrical part 342: Ceiling concave part 343: Second flow path 4: Second plate body 41: Main plate part 41a: First bottom surface , 411: concave portion, 5: adsorption member, 51: block portion, 52: adsorption pad, 53: suction port, 531: extension portion, 532: suction path, 533: projection portion, 54: columnar portion, 541: projection portion , 6: gas permeable membrane, 7: membrane filter, 81, 82, 83: tube, 84: waste liquid outlet, 85: filter, 88: tube, 9: third film member, 92: outer peripheral portion, 93a: bottom surface, 931: recess, 94: port hole, 99: port, C: culture medium, S1: central internal space, S11: first space, S12: second space, S13: third space, S2: surrounding internal space, S4 : fourth space, Sc: sheet-like cells, Hs: heat-sealed portion, x: first direction, x1: first direction one side, x2: first direction other side

Claims (14)

  1. 　第１方向の一方側に位置する第１フィルム状部材、および前記第１フィルム状部材に取り付けられた第１プレート体、を含む第１バッグ部と、
    　前記第１バッグ部に対して前記第１方向の他方側に位置し、外周部が前記第１フィルム状部材の外周部につながる第２フィルム状部材、を含む第２バッグ部と、
    　前記第１プレート体に取り付けられた吸着部材と、を備え、
    　前記吸着部材は、前記第１方向の他方側を向き、且つ前記第２バッグ部に密着可能な環状の吸着パッドを有し、
    　前記第１バッグ部は、各々の一方側端が外部に通じ、且つ他方側端が、前記第１方向に見て前記吸着部材の内側で前記第１方向において前記第１バッグ部と前記第２バッグ部との間の中央内部空間に位置する第１流路および第２流路を有し、
    　前記吸着パッドが前記第２バッグ部に密着する状態にあるとき、前記中央内部空間と、前記第１方向に見て前記吸着部材の外側で前記第１フィルム状部材、前記第２フィルム状部材および前記吸着部材で囲まれた周囲内部空間とが遮断され、
    　前記吸着パッドが前記第２バッグ部に密着しない状態にあるとき、前記中央内部空間と前記周囲内部空間とが連通する、細胞培養用容器。     a first bag portion including a first film-like member located on one side in a first direction and a first plate body attached to the first film-like member;
    a second bag portion including a second film-like member positioned on the other side in the first direction with respect to the first bag portion and having an outer peripheral portion connected to an outer peripheral portion of the first film-like member;
    and an adsorption member attached to the first plate body,
    The suction member has an annular suction pad that faces the other side in the first direction and can be brought into close contact with the second bag portion,
    One side end of each of the first bag portions communicates with the outside, and the other side end of each of the first bag portions and the second bag portion communicates with the first bag portion and the second bag portion in the first direction inside the adsorption member when viewed in the first direction. having a first channel and a second channel located in a central interior space between the bag portion;
    When the suction pad is in close contact with the second bag portion, the first film-like member, the second film-like member, and the second film-like member are formed outside the suction member as viewed in the first direction from the central inner space. The surrounding internal space surrounded by the adsorption member is cut off,
    The cell culture container, wherein the central internal space and the peripheral internal space communicate with each other when the suction pad is not in close contact with the second bag portion.
  2. 　前記第１フィルム状部材は、内周側に第１開口を有する環状とされており、
    　前記第１プレート体は、前記第１開口を塞いでいる、請求項１に記載の細胞培養用容器。     The first film-like member has an annular shape having a first opening on the inner peripheral side,
    The cell culture vessel according to claim 1, wherein said first plate body closes said first opening.
  3. 　前記第２バッグ部は、前記第２フィルム状部材とは別体の第２プレート体を含み、
    　前記第２プレート体は、前記第１方向の一方側を向き、且つ略平坦な第１底面を有し、
    　前記吸着パッドは、前記第１底面に密着可能である、請求項１または２に記載の細胞培養用容器。     The second bag portion includes a second plate body separate from the second film-like member,
    the second plate body faces one side in the first direction and has a substantially flat first bottom surface;
    The cell culture vessel according to claim 1 or 2, wherein the suction pad can be brought into close contact with the first bottom surface.
  4. 　前記第２バッグ部は、前記第２フィルム状部材に取り付けられた第２プレート体を含み、
    　前記第２プレート体は、前記第１方向の一方側を向き、且つ略平坦な第１底面を有し、
    　前記吸着パッドは、前記第１底面に密着可能である、請求項１または２に記載の細胞培養用容器。     The second bag portion includes a second plate body attached to the second film-like member,
    the second plate body faces one side in the first direction and has a substantially flat first bottom surface;
    The cell culture vessel according to claim 1 or 2, wherein the suction pad can be brought into close contact with the first bottom surface.
  5. 　前記第２フィルム状部材は、内周側に第２開口を有する環状とされており、
    　前記第２プレート体は、前記第２開口を塞いでいる、請求項４に記載の細胞培養用容器。     The second film-like member has an annular shape with a second opening on the inner peripheral side,
    Said 2nd plate body is a container for cell cultures of Claim 4 which has block|closed said 2nd opening.
  6. 　前記第２バッグ部は、前記第２フィルム状部材に取り付けられた第２プレート体を含み、
    　前記第２プレート体は、前記第２フィルム状部材に対して前記第１方向の他方側に位置し、且つ前記第１方向に見て前記中央内部空間と重なる、請求項１または２に記載の細胞培養用容器。     The second bag portion includes a second plate body attached to the second film-like member,
    3. The second plate body according to claim 1, wherein said second plate body is positioned on the other side in said first direction with respect to said second film-like member and overlaps said central internal space when viewed in said first direction. Cell culture vessel.
  7. 　前記第１方向に見て、前記第２プレート体の面積は、前記第１プレート体よりも大きい、請求項３ないし６のいずれかに記載の細胞培養用容器。
    7. The cell culture vessel according to any one of claims 3 to 6, wherein said second plate has a larger area than said first plate when viewed in said first direction.
  8. 　前記吸着部材は、前記第１プレート体を前記第１方向に貫通する少なくとも１つの吸引路を有し、
    　前記吸引路は、一方側端が外部に通じており、且つ他方側端が前記吸着パッドの内側に通じる、請求項１ないし７のいずれかに記載の細胞培養用容器。     The adsorption member has at least one suction path penetrating the first plate body in the first direction,
    8. The cell culture vessel according to any one of claims 1 to 7, wherein said suction path communicates with the outside at one end and to the inside of said suction pad at the other end.
  9. 　前記第１フィルム状部材には、各々の一方側端が外部に通じ、且つ他方側端が前記周囲内部空間に通じる複数のポートが設けられている、請求項１ないし８のいずれかに記載の細胞培養用容器。 9. The first film-like member according to any one of claims 1 to 8, wherein the first film-like member is provided with a plurality of ports each of which has one side end communicating with the outside and the other side end communicating with the surrounding inner space. Cell culture vessel.
  10. 　前記第１プレート体は、前記第１方向に見て環状をなす第１環状板部と、前記第１方向に見て前記第１環状板部の内側を塞ぎ、且つ前記第１方向の一方側に張り出す膨出部と、を有し、
    　前記第１環状板部には、ガス透過膜が取り付けられており、
    　前記中央内部空間は、前記第２バッグ部と前記ガス透過膜および前記第１環状板部とに挟まれた第１空間と、前記ガス透過膜および前記膨出部に囲まれた第２空間と、を含み、
    　前記第１プレート体は、各々の一方側端が外部に通じ、且つ他方側端が前記第２空間に通じる複数の第３流路を有する、請求項２に記載の細胞培養用容器。     The first plate body includes a first annular plate portion having an annular shape when viewed in the first direction, a first annular plate portion closing an inner side of the first annular plate portion when viewed in the first direction, and one side of the first direction. and a bulging portion that protrudes into the
    A gas permeable membrane is attached to the first annular plate,
    The central internal space includes a first space sandwiched between the second bag portion, the gas permeable membrane, and the first annular plate portion, and a second space surrounded by the gas permeable membrane and the bulging portion. , including
    3. The container for cell culture according to claim 2, wherein said first plate body has a plurality of third channels, each of which has one side end communicating with the outside and the other side end communicating with said second space.
  11. 　前記第１プレート体には、液体が通過可能な膜フィルターが設けられており、
    　前記中央内部空間は、前記膜フィルターに対して、前記第１方向の他方側に位置する前記第１空間および前記第１方向の一方側に位置する第３空間を含み、
    　前記第１流路の他方側端は、前記第１空間に通じており、
    　前記第２流路の他方側端は、前記第３空間に通じている、請求項１０に記載の細胞培養用容器。     The first plate body is provided with a membrane filter through which liquid can pass,
    the central internal space includes the first space located on the other side in the first direction and the third space located on the one side in the first direction with respect to the membrane filter;
    The other end of the first flow path communicates with the first space,
    11. The cell culture vessel according to claim 10, wherein the other side end of said second channel communicates with said third space.
  12. 　前記第１プレート体のうち前記第３空間を形成する部分には、前記第１方向の一方側に凹む天井凹部が形成されており、
    　前記天井凹部は、前記第２流路につながっている、請求項１１に記載の細胞培養用容器。     A portion of the first plate body that forms the third space is formed with a ceiling recess that is recessed to one side in the first direction,
    The cell culture vessel according to claim 11, wherein the ceiling recess is connected to the second channel.
  13. 　前記第２バッグ部において、前記第１方向の一方側を向く面のうち前記第１方向に見て前記吸着パッドの内側を含む領域には、稠密に配列された複数の凹部が設けられている、請求項１ないし１２のいずれかに記載の細胞培養用容器。 In the second bag portion, a plurality of densely arranged recesses are provided in a region of the surface facing one side in the first direction that includes the inner side of the suction pad when viewed in the first direction. 13. The container for cell culture according to any one of claims 1 to 12.
  14. 　外周部が前記第２フィルム状部材の前記外周部につながる第３フィルム状部材をさらに備え、
    　前記第２バッグ部の前記第１方向の他方側と前記第３フィルム状部材とに挟まれた第４空間を有し、
    　前記吸着パッドが前記第２バッグ部に密着しない状態にあるとき、前記中央内部空間と前記第４空間とが連通する連通手段を備える、請求項１ないし１３のいずれかに記載の細胞培養用容器。     further comprising a third film-like member having an outer peripheral portion connected to the outer peripheral portion of the second film-like member;
    a fourth space sandwiched between the other side of the second bag portion in the first direction and the third film-like member;
    14. The container for cell culture according to any one of claims 1 to 13, further comprising communicating means for communicating between said central internal space and said fourth space when said suction pad is not in close contact with said second bag portion. .
PCT/JP2021/048882 2021-12-28 2021-12-28 Container for cell culture use WO2023127132A1 (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021000017A (en) * 2019-06-20 2021-01-07 富士フイルム株式会社 Flow path forming member
WO2021014642A1 (en) * 2019-07-25 2021-01-28 株式会社サンプラテック Container for housing culture solution

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021000017A (en) * 2019-06-20 2021-01-07 富士フイルム株式会社 Flow path forming member
WO2021014642A1 (en) * 2019-07-25 2021-01-28 株式会社サンプラテック Container for housing culture solution

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