Cell culture container

Abstract

This cell culture container comprises: a container member including a cylindrical part in which a culture space for accommodating a plurality of cells is formed; and a bottom sheet joined to the container member to close one end of the cylindrical part. The bottom sheet is a sheet formed of an elastic material. The bottom sheet includes a joining surface joined to an end surface of the cylindrical part, and a projecting part projecting from the joining surface and coming into contact with the inner wall surface of the cylindrical part.

Description

cell culture container

[0001] The present invention relates to a cell culture vessel used for culturing cells.

[0002] Cell culture vessels (well plates) for culturing various types of cells have been proposed. For example, Patent Document 1 discloses a biochemical plate having a plurality of cylindrical bodies attached to the surface of a bottom plate. The bottom plate is made of glass. Protrusions are attached to the surface of the bottom plate, and each cylindrical body is fitted onto the protrusions.

[0003] Japanese Patent Application Laid-Open No. 2006-112917

[0004] Dimensional errors may occur in the bottom plate and the multiple cylindrical bodies. Because the bottom plate of Patent Document 1 is a hard substrate made of glass, if there are dimensional errors in the bottom plate and the multiple cylindrical bodies, it is difficult to fit all of the multiple cylindrical bodies onto the respective protrusions of the bottom plate. In consideration of the above circumstances, one aspect of the present disclosure aims to easily position a specific area of ​​the bottom sheet inside the cylindrical portion while absorbing dimensional errors between the container member including the cylindrical portion and the bottom sheet.

[0005] A cell culture container according to one aspect of the present disclosure comprises a container member including a tubular portion having a culture space formed therein for accommodating a plurality of cells, and a bottom sheet that is joined to the container member to close one end of the tubular portion, the bottom sheet being a sheet formed of an elastic material, and including a joining surface that is joined to the end face of the tubular portion, and a protrusion that protrudes from the joining surface and contacts the inner wall surface of the tubular portion.

[0006] 1 is an exploded perspective view of a cell culture vessel according to an embodiment. FIG. 1 is an exploded perspective view of a cell culture vessel according to an embodiment. FIG. 2 is a plan view focusing on one culture space. FIG. 3 is a cross-sectional view taken along line IV-IV in FIG. 3. FIG. 4 is a cross-sectional view of a comparative example. FIG. 5 is an explanatory view of a problem in the comparative example. FIG. 6 is an explanatory view of application of a bonding material to a bottom sheet 20. FIG. 7 is a cross-sectional view of a cell culture vessel according to a second embodiment. FIG. 8 is a cross-sectional view of a cell culture vessel according to a modified example of the first embodiment. FIG. 9 is a cross-sectional view of a cell culture vessel according to a modified example of the first embodiment. FIG. 10 is a cross-sectional view of a cell culture vessel according to a modified example of the first embodiment. FIG. 11 is a cross-sectional view of a cell culture vessel according to a modified example of the first embodiment. FIG. 12 is a cross-sectional view of a cell culture vessel according to a modified example of the first embodiment. FIG. 13 is a cross-sectional view of a cell culture vessel according to a modified example of the second embodiment. FIG. 14 is a cross-sectional view of a cell culture vessel according to a modified example of the second embodiment. FIG. 15 is a cross-sectional view of a cell culture vessel according to a modified example of the second embodiment. FIG. 16 is a cross-sectional view of a cell culture vessel according to a modified example.

[0007] The embodiments for carrying out the present disclosure will be described with reference to the drawings. Note that the dimensions and scale of each element in each drawing may differ from those of the actual product. Furthermore, the embodiment described below is an exemplary embodiment that may be envisioned when carrying out the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiment exemplified below.

[0008] 1 and 2 are exploded perspective views of a cell culture vessel 100 according to one embodiment of the present disclosure. The cell culture vessel 100 is a vessel (well plate) used for culturing various types of cells.

[0009] In the following description, the Z axis is assumed. One direction along the Z axis is referred to as the Z1 direction, and the opposite direction to the Z1 direction is referred to as the Z2 direction. The Z axis is, for example, an axis along the vertical direction. The Z1 direction is a downward vertical direction, and the Z2 direction is an upward vertical direction. However, the orientation of the cell culture vessel 100 in actual use is arbitrary. In the following description, observing each element of the cell culture vessel 100 from a line of sight along the Z axis is referred to as a "planar view."

[0010] As illustrated in FIGS. 1 and 2, the cell culture vessel 100 comprises a vessel member 10 and a bottom sheet 20 .

[0011] The container member 10 is a flat structure formed in a generally rectangular shape. The container member 10 is made of various resin materials such as polystyrene, polypropylene, polycarbonate, or polyester. The container member 10 may also be made of a combination of multiple members made of different materials.

[0012] 1 and 2 , the container member 10 includes a side wall portion 11, a plurality of cylindrical portions 12, and a base portion 13. The side wall portion 11 is a rectangular frame-shaped portion that forms the side surface of the container member 10. The plurality of cylindrical portions 12 and the base portion 13 are installed inside the side wall portion 11.

[0013] The multiple cylindrical portions 12 are arranged in a matrix along a plane (e.g., a horizontal plane) perpendicular to the Z axis. The cylindrical portions 12 are installed at intervals from each other. The number of cylindrical portions 12 is arbitrary, but for example, a total of 96 cylindrical portions 12, consisting of 8 rows and 12 columns, are arranged inside the side wall portion 11. Each of the multiple cylindrical portions 12 is a cylindrical portion whose central axis is parallel to the Z axis. A culture space S for accommodating cells is formed inside each cylindrical portion 12. That is, the cell culture vessel 100 of the first embodiment is a vessel in which multiple culture spaces S are installed.

[0014] 2, the base portion 13 is a plate-like portion that closes the gaps between the cylindrical portions 12 inside the side wall portion 11. The back surface 14 of the container member 10 is composed of the end faces of the cylindrical portions 12 facing the Z1 direction and the main surface (i.e., the bottom surface) of the base portion 13 facing the Z1 direction. Circular openings corresponding to the cylindrical portions 12 are formed in the back surface 14 of the container member 10.

[0015] The bottom sheet 20 is a sheet-like member that is more flexible than the container member 10 and is formed into the same external shape as the container member 10. The bottom sheet 20 is formed from various elastic materials and is elastically stretchable. The bottom sheet 20 is also formed from an oxygen-permeable transparent resin material. Examples of materials for the bottom sheet 20 include silicone elastomers such as polydimethylsiloxane (PDMS). The bottom sheet 20 may also be formed by laminating multiple layers made of different materials. The thickness of the bottom sheet 20 is, for example, 3 mm or less (more preferably 2 mm or less).

[0016] The cell culture vessel 100 is constructed by bonding the bottom sheet 20 to the container member 10. Specifically, the surface of the bottom sheet 20 facing the container member 10 (hereinafter referred to as the "facing surface 21") is bonded to the back surface 14 of the container member 10. By bonding the bottom sheet 20 to the back surface 14 of the container member 10, the opening at one end of each tubular portion 12 is closed. A bonding material such as a silane coupling agent is used to bond the container member 10 and the bottom sheet 20. Note that the container member 10 and the bottom sheet 20 may also be bonded using an adhesive formed from various resin materials.

[0017] Fig. 3 is a plan view focusing on one arbitrary culture space S. Fig. 4 is a cross-sectional view taken along line IV-IV in Fig. 3. The configuration illustrated in Figs. 3 and 4 for one culture space S is common to the multiple culture spaces S of the cell culture vessel 100. In the following description, the direction of the radius of a virtual circle of an arbitrary diameter concentric with the cylindrical portion 12 in a plan view is referred to as the "radial direction." Furthermore, the direction toward the center of the cylindrical portion 12 in the radial direction is referred to as the "inner side."

[0018] 4 , the opposing surface 21 of the bottom sheet 20 includes a bonding surface 22 that is bonded to the back surface 14 of the container member 10. The bonding surface 22 is the area of ​​the opposing surface 21 of the bottom sheet 20 that faces the back surface 14 of the container member 10. As described above, the back surface 14 includes the end surface of the tubular portion 12, and therefore the bonding surface 22 can also be expressed as the area of ​​the surface of the bottom sheet 20 that is bonded to the end surface of the tubular portion 12. An opening at one end of the tubular portion 12 in the Z1 direction is closed by the bottom sheet 20, thereby forming a culture space S inside the tubular portion 12. In other words, the culture space S is a cylindrical space surrounded by the inner wall surface 15 of the tubular portion 12.

[0019] A protrusion 23 and a bottom surface 24 are formed on the opposing surface 21 of the bottom sheet 20. As illustrated in FIG.

[0020] The protrusion 23 is a portion that protrudes in the Z direction from the joining surface 22. As illustrated in Figures 3 and 4, the protrusion 23 of the first embodiment is formed in an annular shape along the inner wall surface 15 of the tubular portion 12 in a plan view.

[0021] Specifically, the protrusion 23 includes an outer wall surface 231 and an inclined surface 232. The outer wall surface 231 is a surface of revolution that protrudes in the Z direction from the joining surface 22. Specifically, the outer wall surface 231 is a cylindrical surface that constitutes the outer peripheral edge Eb of the protrusion 23.

[0022] The outer diameter of the protrusion 23 (i.e., the diameter of the outer wall surface 231) is set to a dimension equal to the inner diameter of the inner wall surface 15 or a dimension slightly larger than the inner diameter of the inner wall surface 15. Therefore, the outer wall surface 231 contacts the inner wall surface 15 of the tubular portion 12. Specifically, the outer wall surface 231 is in close contact with the inner wall surface 15 over the entire circumference. As described above, the protrusion 23 contacts the inner wall surface 15 of the tubular portion 12. In other words, the protrusion 23 is fitted into the tubular portion 12.

[0023] The inclined surface 232 is a surface of revolution extending inward from the upper edge U (outer peripheral edge Eb) of the outer wall surface 231 to the inner peripheral edge Ea of the protrusion 23. The angle of elevation between the outer wall surface 231 and the inclined surface 232 is an acute angle. Specifically, the inclined surface 232 is a truncated cone surface in which the inner peripheral edge Ea is lower than the outer peripheral edge Eb. That is, the inclined surface 232 is inclined with respect to the Z axis so as to descend from the outer peripheral edge Eb toward the inner peripheral edge Ea. Therefore, the position of the upper edge U (outer peripheral edge Eb) on the Z axis is located in the Z2 direction relative to the position of the inner peripheral edge Ea on the Z axis. The inclined surface 232 can also be expressed as a tapered surface whose generatrix is ​​a straight line.

[0024] The bottom surface 24 is a region of the opposing surface 21 of the bottom sheet 20 that is located radially inside the protrusion 23 in a plan view. That is, the bottom surface 24 is a circular plane that continues to the inner peripheral edge Ea of the protrusion 23 and is surrounded by the protrusion 23. Therefore, the bottom surface 24 is located lower than the upper edge U of the outer wall surface 231 of the protrusion 23. Specifically, the bottom surface 24 is a plane that is at the same height as the joint surface 22. Therefore, the protrusion 23 can also be expressed as a portion that protrudes from the bottom surface 24 in the Z2 direction. The culture space S is a space surrounded by the inner wall surface 15 of the tubular portion 12 above the inclined surface 232 and the bottom surface 24 (in the Z2 direction).

[0025] A plurality of recesses 25 are formed on the bottom surface 24. As illustrated in FIG. 1, a group of a plurality of recesses 25 is formed on the bottom sheet 20, spaced apart from one another, for each culture space S. Each recess 25 is a bottomed hole (dimple) that accommodates cells. In the first embodiment, each recess 25 is, for example, a hemispherical or cylindrical depression. However, the shape of the recess 25 is not limited to the above examples.

[0026] The hole diameter (outer diameter) Wa of the recess 25 is, for example, 0.5 mm. The hole diameter Wa of the recess 25 is, for example, a numerical value obtained by averaging the outer diameters of the recesses 25 over a plurality of recesses 25 corresponding to one culture space S. The depth of the recess 25 is, for example, 1.6 mm. The depth of the recess 25 is the distance between the bonding surface 22 and the bottom of the recess 25.

[0027] The width Wb of the protrusion 23 is greater than the hole diameter Wa of each of the recesses 25 (Wb > Wa). The width Wb of the protrusion 23 is the distance between the inner peripheral edge Ea and the outer peripheral edge Eb in a plan view. More specifically, the width Wb is, for example, a value obtained by averaging the width of the protrusion 23 at each point in the circumferential direction over the entire circumference of the protrusion 23. As described above, the protrusion 23 is formed wider than the recesses 25. The width Wb of the protrusion 23 is set to a value within the range of 0.5 mm or more and 3 mm or less, for example.

[0028] A culture solution in which a large number of cells are dispersed is supplied to the culture space S. The large number of cells seeded in the culture space S are accommodated in each of the plurality of recesses 25. Within each recess 25, the plurality of cells aggregate to form a three-dimensional cell mass (spheroid). The inner surface of the culture space S (the inclined surface 232, the bottom surface 24, and the inner wall surface 15) may be coated with, for example, a resin material.

[0029] As described above, the bottom sheet 20 is made of an oxygen-permeable material, so that oxygen is sufficiently supplied to the cell clusters in the culture space S. Therefore, necrosis of the cell clusters due to a lack of oxygen is suppressed. As described above, the survival rate of the cell clusters is maintained at a high level, and as a result, high-quality cell clusters can be formed. In particular, in this embodiment, the bottom sheet 20 is made of polydimethylsiloxane, so that the bottom sheet 20 is transparent, making it easy to observe the cells, and is also advantageous in that it is non-toxic to cells and makes it easy to form fine structures.

[0030] As described above, the protrusion 23 of the first embodiment includes an inclined surface 232 in which the inner periphery Ea is lower than the outer periphery Eb. In the above configuration, among the many cells seeded in the culture space S, cells that reach the vicinity of the inner wall surface 15 move along the inclined surface 232 toward the inner periphery Ea of the protrusion 23, as shown by the dashed arrow in FIG. 4 . In other words, the cells can be guided to the center of the culture space S. Specifically, in the first embodiment, among the many cells seeded in the culture space S, cells located near the inner wall surface 15 move along the inclined surface 232 and are accommodated in the multiple recesses 25 on the bottom surface 24. In other words, a sufficient number of cells can be accommodated in the multiple recesses 25.

[0031] 4 is the distance between the upper edge U of the outer wall surface 231 and the joint surface 22. The height H is set to a value within the range of 1 mm or more and 5 mm or less, for example.

[0032] Incidentally, in a configuration in which the width Wb of the protrusion 23 is excessively large relative to the height H, the inclination of the inclined surface 232 is sufficiently small, so there is a possibility that cells will remain on the inclined surface 232. In other words, there is a possibility that cells present near the inner wall surface 15 will not be guided to the center of the culture space S. On the other hand, in a configuration in which the height H of the protrusion 23 is excessively large relative to the width Wb, the protrusion 23 is easily deformed by pressure from the inner wall surface 15 of the cylindrical portion 12.

[0033] The width Wb and height H of the protrusion 23 are set to appropriate dimensions so as to prevent cells from remaining on the inclined surface 232 and to prevent excessive deformation of the protrusion 23, as described above. For example, the ratio of the height H to the width Wb of the protrusion 23 (H / Wb) is set to an appropriate value within the range of 0.25 or more and 2 or less. As can be understood from the above explanation, possible configurations are those in which the width Wb of the protrusion 23 exceeds the height H, those in which the width Wb of the protrusion 23 is less than the height H, and those in which the width Wb and height H of the protrusion 23 are equal to each other.

[0034] As described above, in the first embodiment, the protrusions 23 are formed on the bottom sheet 20 made of an elastic material. Therefore, the bottom sheet 20 expands and contracts to absorb dimensional errors between the container member 10 and the bottom sheet 20, and the protrusions 23 contact the inner wall surface 15 of the tubular portion 12, making it possible to easily position the bottom surface 24 inside the tubular portion 12. In other words, there is an advantage in that joining the bottom sheet 20 to the container member 10 is made easier. As can be understood from the above description, each protrusion 23 functions as an element for positioning each bottom surface 24 of the bottom sheet 20 inside the tubular portion 12.

[0035] Furthermore, in the first embodiment, since the plurality of recesses 25 are formed inside the protrusion 23, there is an advantage that the plurality of recesses 25 are less affected by the fit between the cylindrical portion 12 and the protrusion 23 (i.e., the positioning of the bottom surface 24). Specifically, the possibility that each recess 25 will be deformed by stress caused by the fit of the protrusion 23 to the cylindrical portion 12 is reduced.

[0036] In the first embodiment, in particular, the outer wall surface 231 of the annularly formed protrusion 23 comes into contact with the inner wall surface 15 of the tubular portion 12. Therefore, the position of the bottom sheet 20 relative to the container member 10 can be determined in all directions within the plane of the bottom sheet 20.

[0037] FIG. 5 is a cross-sectional view of a configuration (hereinafter referred to as the "Comparative Example") in which the protrusions 23 are not formed. In the Comparative Example, because there is no structure (the protrusions 23 of the first embodiment) for positioning the bottom surface 24 relative to the tubular portion 12, an error may occur in the position of the bottom surface 24 relative to the tubular portion 12. If an error occurs in the position of the bottom surface 24, as illustrated in FIG. 6 , some of the multiple recesses 25 may overlap the tubular portion 12 in a plan view, resulting in the occlusion of the recesses 25. In contrast to the Comparative Example, in the first embodiment, the protrusions 23 contact the inner wall surface 15 of the tubular portion 12, making it possible to easily position the bottom surface 24 inside the tubular portion 12. Therefore, according to the first embodiment, the occlusion of the recesses 25 can be suppressed.

[0038] As described above, the bottom sheet 20 is bonded to the rear surface 14 of the container member 10 with a bonding material such as a silane coupling agent or a resin material. In a comparative example in which the protrusions 23 are not formed, it is practically difficult to selectively apply the bonding material only to the bonding surface 22 of the opposing surface 21 of the bottom sheet 20, and the bonding material may also adhere to the bottom surface 24. However, if the bonding material adheres to the bottom surface 24, it may cause problems with coating the bottom surface 24 or cell culture.

[0039] In contrast to the comparative example, in the first embodiment, the bottom surface 24 is surrounded by the protrusions 23 protruding from the bonding surface 22, so that it is possible to apply the bonding material with high precision only to the bonding surface 22 of the opposing surface 21 of the bottom sheet 20, as illustrated in Fig. 7. In other words, adhesion of the bonding material to the bottom surface 24 is suppressed. Therefore, according to the first embodiment, there is an advantage that various problems caused by adhesion of the bonding material to the bottom surface 24 can be reduced compared to the comparative example.

[0040] Furthermore, in the first embodiment, the width Wb of the protrusion 23 is greater than the hole diameter Wa of each of the plurality of recesses 25. According to the above configuration, the mechanical strength of the protrusion 23 is ensured compared to a configuration in which the width Wb of the protrusion 23 is smaller than the hole diameter Wa of the recess 25, and therefore deformation of the protrusion 23 due to pressure from the inner wall surface 15 of the tubular portion 12 can be suppressed.

[0041] B: Second Embodiment A second embodiment will be described. Note that, for elements in the following exemplary aspects that have the same functions as those in the first embodiment, the same reference numerals as those in the first embodiment will be used, and detailed descriptions of each element will be omitted as appropriate.

[0042] 8 is a cross-sectional view focusing on one culture space S of the cell culture vessel 100 according to the second embodiment. As illustrated in FIG. 8, a protrusion 23 and a bottom surface 24 are formed on the opposing surface 21 of the bottom sheet 20.

[0043] The protrusion 23 is a portion that protrudes in the Z2 direction from the joint surface 22 and is formed in a ring shape that follows the inner wall surface 15 of the tubular portion 12 in a plan view. The protrusion 23 in the second embodiment includes an outer wall surface 231. As in the first embodiment, the outer wall surface 231 is a surface of revolution that protrudes in the Z2 direction from the joint surface 22. Specifically, the outer wall surface 231 is a cylindrical surface that constitutes the outer peripheral edge Eb of the protrusion 23.

[0044] The bottom surface 24 of the second embodiment is a circular region of the opposing surface 21 of the bottom sheet 20 that is located inside the outer wall surface 231 in a plan view. The bottom surface 24 is a concave surface with a central portion C that is lower than the outer peripheral edge Eb. Specifically, the bottom surface 24 is a spherical surface that is recessed with respect to the upper edge U of the outer wall surface 231. The central portion C of the bottom surface 24 is located lower than the joining surface 22.

[0045] The culture space S in the second embodiment is a space surrounded by the inner wall surface 15 of the cylindrical portion 12 above the bottom surface 24 (in the Z2 direction). Note that the bottom surface 24 in the second embodiment does not have multiple recesses 25 formed therein. Therefore, cell aggregates are formed near the center C of the bottom surface 24. Note that the inner surface of the culture space S (bottom surface 24 and inner wall surface 15) may be coated with, for example, a resin material.

[0046] The second embodiment also achieves the same effects as the first embodiment. Furthermore, in the second embodiment, among the many cells seeded in the culture space S, cells located near the inner wall surface 15 migrate along the concave surface of the bottom surface 24 to the center C. That is, the cells can be guided to the center C of the bottom surface 24 of the culture space S.

[0047] C: Modifications Specific modifications that can be added to the above-mentioned embodiments are exemplified below. Two or more embodiments arbitrarily selected from the following examples may be combined as appropriate within the scope of not mutually contradicting each other.

[0048] (1) In the first embodiment, the bottom surface 24 is positioned at the same height as the joining surface 22, but the height of the bottom surface 24 is not limited to the above example. For example, a configuration in which the bottom surface 24 is positioned higher than the joining surface 22 as illustrated in Fig. 9 or a configuration in which the bottom surface 24 is positioned lower than the joining surface 22 as illustrated in Fig. 10 is also possible.

[0049] (2) The shape of the protrusion 23 is not limited to the example shown in the first embodiment. For example, in the first embodiment, the inclined surface 232 of the protrusion 23 is a truncated cone surface (tapered surface), but the shape of the inclined surface 232 is not limited to a truncated cone surface. For example, as shown in FIG. 11 , the inclined surface 232 may be a convex arc surface facing the Z2 direction, or as shown in FIG. 12 , the inclined surface 232 may be a concave arc surface facing the Z2 direction. Note that the shape of the generatrix of the inclined surface 232 is not limited to the above example. For example, the inclined surface 232 may be a surface of revolution whose generatrix is ​​a parabolic curve, a cycloid curve, or any other curve.

[0050] Furthermore, in the first embodiment, the cross-sectional shape of the protrusion 23 is exemplified as a triangle including the inclined surface 232, but as exemplified in Fig. 13, the cross-sectional shape of the protrusion 23 may be rectangular. That is, the inclined surface 232 may be omitted. However, in the embodiment of Fig. 13, among the large number of cells seeded in the culture space S, cells that reach the vicinity of the inner wall surface 15 may remain on the upper surface of the protrusion 23. Therefore, from the viewpoint of efficiently accommodating the large number of cells seeded in the culture space S in the multiple recesses 25, the embodiment in which the protrusion 23 includes the inclined surface 232 (Figs. 4, 11, and 12) is preferable.

[0051] (3) In the second embodiment, the central portion C of the bottom surface 24 is located at a position lower than the joining surface 22. However, the position of the central portion C in the Z-axis direction is not limited to the above example. For example, a configuration in which the central portion C is at the same height as the joining surface 22 as illustrated in Fig. 14 or a configuration in which the central portion C is located at a position higher than the joining surface 22 as illustrated in Fig. 15 is also possible.

[0052] (4) In the second embodiment, the bottom surface 24 is illustrated as a spherical surface, but the shape of the bottom surface 24 is not limited to this example. For example, the bottom surface 24 may be a conical surface as illustrated in FIG. 16 or a truncated conical surface as illustrated in FIG. 17 . The configuration illustrated in FIG. 17 can also be expressed as a configuration in which the multiple recesses 25 in the first embodiment are omitted. As described above with reference to FIGS. 14 and 15 , the position of the center C of the bottom surface 24 is arbitrary in the configurations of FIGS. 16 and 17 .

[0053] (5) In the above-described embodiments, the bottom surface 24 is located at a position lower than the upper edge U of the outer wall surface 231 of the protrusion 23. However, as illustrated in FIG. 18 , a configuration in which the bottom surface 24 is located at the same height as the upper edge U of the outer wall surface 231 is also possible. However, in the configuration of FIG. 18 , the outer wall surface 231 of the protrusion 23 is pressed by the inner wall surface 15 of the cylindrical portion 12, which easily deforms the bottom surface 24. Deformation of the bottom surface 24 may cause distortion in the recess 25. According to the configuration in which the bottom surface 24 is located at a position lower than the upper edge U of the outer wall surface 231, deformation of the bottom surface 24 due to pressure from the inner wall surface 15 of the cylindrical portion 12 is suppressed. This has the advantage of reducing distortion in the recess 25.

[0054] (6) The dimensions of each element of the cell culture vessel 100 are not limited to the above examples. For example, in the first embodiment, the width Wb of the protrusion 23 is greater than the hole diameter Wa of the recess 25, but a configuration in which the width Wb of the protrusion 23 and the hole diameter Wa of the recess 25 are equal is also conceivable.

[0055] 19 and 20, a configuration in which the width Wb of the protrusion 23 is smaller than the hole diameter Wa of the recess 25 (Wb<Wa) is also possible. In the configuration of Fig. 19, a plurality of recesses 25 are formed in the bottom surface 24 up to an area near the inner wall surface 15 of the cylindrical portion 12. In the configuration of Fig. 20, recesses 25 with larger diameters than those in the first embodiment are formed in the bottom surface 24.

[0056] 19 and 20, in which the width Wb of the protrusion 23 is smaller than the hole diameter Wa of the recess 25, the bottom surface 24 is enlarged compared to a configuration in which the width Wb exceeds the hole diameter Wa (for example, the first embodiment), making it easier to increase the number of recesses 25 (FIG. 19) or increase the hole diameter Wa of the recess 25 (FIG. 20). Therefore, there is an advantage in that a larger number of cell aggregates can be cultured even if the seeding amount in the culture space S is the same.

[0057] In addition, in the first embodiment, a configuration in which the height H of the protrusion 23 is less than the width Wb of the protrusion 23 is exemplified, but a configuration in which the height H of the protrusion 23 exceeds the width Wb of the protrusion 23, or a configuration in which the height H and the width Wb are equal, is also envisioned.

[0058] (7) In the above embodiment, the protrusion 23 is formed in an annular shape, but the planar shape of the protrusion 23 is not limited to the above example. For example, as shown in Fig. 21, the protrusion 23 may be formed of a plurality of portions 23a. The plurality of portions 23a shown in Fig. 21 are arc-shaped portions arranged at intervals in the circumferential direction.

[0059] (8) In each of the above embodiments, a well plate is exemplified as the cell culture vessel 100, but the vessel to which the present disclosure is applicable is not limited to the above examples. For example, the present disclosure is applicable to any type of cell culture vessel, such as a petri dish or a culture flask.

[0060] D: Supplementary Notes From the above-described exemplary embodiments, the following configurations can be understood, for example.

[0061] A cell culture vessel according to one aspect (Aspect 1) of the present disclosure includes a container member including a cylindrical portion having a culture space formed therein for accommodating a plurality of cells, and a bottom sheet joined to the container member to close one end of the cylindrical portion. The bottom sheet is made of an elastic material and includes a joining surface joined to the end face of the cylindrical portion and a protrusion protruding from the joining surface and contacting the inner wall surface of the cylindrical portion. In the above aspect, the protrusion is formed on the bottom sheet made of an elastic material, and the protrusion contacts the inner wall surface of the cylindrical portion. Therefore, the bottom surface of the culture space can be easily positioned inside the cylindrical portion by contacting the protrusion with the inner wall surface of the cylindrical portion while absorbing dimensional errors between the container member and the bottom sheet by expanding and contracting the bottom sheet.

[0062] In a specific example (Aspect 2) of Aspect 1, the protrusion is formed in a ring shape along the inner wall surface, and the outer wall surface of the protrusion contacts the inner wall surface of the cylindrical portion. In the above aspect, the outer wall surface of the protrusion formed in a ring shape contacts the inner wall surface of the cylindrical portion. Therefore, the position of the bottom sheet relative to the container member can be determined in all directions within the plane of the bottom sheet.

[0063] In a specific example of Aspect 2 (Aspect 3), the protrusions include an inclined surface whose inner periphery is lower than the outer periphery. In this aspect, among the many cells seeded in the culture space, cells that reach the vicinity of the inner wall surface migrate along the inclined surface toward the inner periphery of the protrusions. In other words, the cells can be guided to the center of the culture space.

[0064] In a specific example (Aspect 4) of Aspect 3, the bottom sheet includes a bottom surface located radially inside the protrusions, and a plurality of recesses for accommodating the plurality of cells are formed on the bottom surface. In the above aspect, among the large number of cells seeded in the culture space, cells located near the inner wall surface are accommodated in the plurality of recesses by moving along the inclined surface. Therefore, a sufficient number of cells can be accommodated in the plurality of recesses.

[0065] In a specific example (Aspect 5) of Aspect 4, the width of the protrusion is greater than the diameter of each of the recesses. According to this aspect, the mechanical strength of the protrusion is ensured, and deformation of the protrusion due to pressure from the inner wall surface can be suppressed.

[0066] In a specific example (Aspect 6) of Aspect 4, the width of the protrusions is less than the diameter of each of the plurality of wells. This aspect makes it easier to ensure a sufficient bottom surface area compared to an embodiment in which the width of the protrusions exceeds the diameter of the wells. This has the advantage of allowing a large number of cell aggregates to be cultured.

[0067] In a specific example (Aspect 7) of any one of Aspects 3 to 6, the ratio of the height of the protrusions to the width of the protrusions is not less than 0.25 and not more than 2. According to the above aspect, it is possible to suppress excessive deformation of the protrusions due to pressure from the inner wall surface of the tubular portion while suppressing cells from remaining on the inclined surface of the protrusions.

[0068] A specific example of Aspect 2 (Aspect 8) includes a bottom surface located radially inside the outer wall surface, and the bottom surface is a concave surface with a central portion lower than the outer periphery. In this aspect, among the many cells seeded in the culture space, cells located near the inner wall surface migrate to the central portion along the concave surface of the bottom. In other words, the cells can be guided to the central portion of the bottom of the culture space.

[0069] 100...cell culture vessel, 10...vessel member, 11...side wall portion, 12...cylindrical portion, 13...base portion, 14...back surface, 15...inner wall surface, 20...bottom sheet, 21...opposing surface, 22...joint surface, 23...protrusion portion, 231...outer wall surface, 232...inclined surface, 24...bottom surface, 25...recess, S...culture space.

Claims

1. A cell culture vessel comprising: a container member including a cylindrical portion in which a culture space for accommodating a plurality of cells is formed; and a bottom sheet that is joined to the container member to close one end of the cylindrical portion, the bottom sheet being a sheet made of an elastic material, the bottom sheet including a joining surface that is joined to the end face of the cylindrical portion, and a protrusion that protrudes from the joining surface and contacts the inner wall surface of the cylindrical portion.

2. The cell culture vessel according to claim 1, wherein the protrusion is formed in a ring shape along the inner wall surface, and the outer wall surface of the protrusion contacts the inner wall surface of the cylindrical portion.

3. The cell culture vessel according to claim 2, wherein the protrusion includes an inclined surface whose inner periphery is lower than its outer periphery.

4. The cell culture vessel according to claim 3, wherein the bottom sheet includes a bottom surface positioned radially inside the protrusions, and a plurality of recesses for accommodating the plurality of cells are formed on the bottom surface.

5. The cell culture vessel according to claim 4, wherein the width of the protrusion is greater than the diameter of each of the plurality of recesses.

6. The cell culture vessel according to claim 4, wherein the width of the protrusion is less than the diameter of each of the plurality of recesses.

7. A cell culture vessel according to any one of claims 3 to 6, wherein the ratio of the height of the protrusion to the width of the protrusion is 0.25 or more and 2 or less.

8. The cell culture vessel according to claim 2, further comprising a bottom surface located radially inside the outer wall surface, the bottom surface being a concave surface with a central portion lower than the outer periphery.

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