Culture vessel and method for producing culture vessel

Abstract

This culture vessel comprises: a body formed into a cylinder shape and having an opening at the top end; and a base member anchored to the bottom end of the body to form the base of the body, the base member having a culture surface formed thereon, on which cells can be cultured. The base member is formed from a degradable material.

Description

Culture vessel and method for manufacturing the culture vessel

[0001] The technology of the present disclosure relates to a culture vessel and a method for manufacturing the culture vessel.

[0002] JP 2011-517696 A describes a non-biodegradable membrane for supporting cell growth. According to this document, the membrane on which the cells have been cultured is cut with a scalpel in a tissue culture flask, and the cells are recovered. Furthermore, JP 2005-348736 A describes a support device for a cell culture membrane for culturing corneal cells. According to this document, the cell culture membrane on which the corneal cells have been cultured is sandwiched and held between upper and lower members.

[0003] In the technique described in JP-A-2011-517696, cells are cultured on a non-biodegradable membrane, which limits its applications, whereas in the technique described in JP-A-2005-348736, a degradable substrate is used for the cell culture membrane, which can be decomposed in vivo, for example.

[0004] Here, when harvesting cell fragments cultured on a membrane body or the like, it is desirable to simplify the process as much as possible. However, the technique disclosed in JP 2011-517696 A requires the use of a scalpel to cut the membrane, which requires the skilled technical skills of the worker. Furthermore, although JP 2005-348736 A describes harvesting corneas of the desired size by punching, the cell culture membrane is held between upper and lower members, which may require the time-consuming task of disassembling the upper and lower members to remove the cell culture membrane when harvesting the cornea.

[0005] The technology according to the present disclosure provides a culture vessel and a culture method that can reduce the effort required for collecting cells.

[0006] A first aspect of the culture vessel comprises a cylindrical main body portion having an opening at the upper end, and a bottom member that is fixed to the lower end of the main body portion to form the bottom of the main body portion and has a culture surface on which cells can be cultured, the bottom member being formed from a degradable material.

[0007] In the culture vessel of the first embodiment, cells can be cultured on the bottom member. The bottom member is fixed to the lower end of the main body and forms the bottom of the main body. Therefore, by placing the culture vessel on a base or the like, inserting a cell collection tool from the upper end of the cylindrical main body, and pressing the tip of the cell collection tool against the bottom member, a reaction force is generated from the base, allowing the cultured cells to be punched out together with the bottom member and collected.

[0008] Thus, compared to a configuration in which the cell culture membrane is fixed by sandwiching it between multiple components from above and below, this culture vessel eliminates the need to disassemble the components when harvesting cells. In other words, the labor required for harvesting cells can be reduced. Furthermore, because the number of components can be reduced, there is less risk of contamination.

[0009] Furthermore, the bottom member is made of a degradable material, so that after cells are collected together with the bottom member using a cell collection instrument, the bottom member can be decomposed and the cells alone can be used (for example, for biotransplantation).

[0010] A culture vessel of a second aspect is the culture vessel of the first aspect, wherein the bottom member is bonded to the lower end surface of the main body portion.

[0011] In the culture vessel of the second aspect, the bottom member is formed separately from the main body and is bonded to the main body, which allows selection of a material suitable for the performance required of the bottom member.

[0012] A culture vessel of a third aspect is the culture vessel of the second aspect, wherein the bottom member is heat-welded to the lower end surface.

[0013] In the culture vessel of the third aspect, the bottom member is heat-welded to the main body, which eliminates the need to use materials such as adhesives.

[0014] In a fourth aspect of the culture vessel, in the culture vessel of the third aspect, the bottom member is heat-welded to a portion of the lower end surface that extends from a position offset outward from the culture surface to the outer side.

[0015] In the culture vessel of the fourth aspect, the bottom member is heat-welded to the outer portion of the lower end surface of the main body from a position offset outward from the culture surface, thereby reducing the thermal influence on the culture surface compared to when there is no offset.

[0016] The culture vessel of the fifth embodiment is the culture vessel of the third embodiment, wherein the surface free energy of the main body is −10 mJ / m 2 or more and +10mJ / m 2 The following is the result.

[0017] According to the culture vessel of the fifth aspect, the surface free energy of the main body is −10 mJ / m 2 If it is smaller than this, it will be +10mJ / m 2 Compared with a larger diameter, the bondability by thermal welding is improved.

[0018] A culture vessel of a sixth aspect is the culture vessel of the first aspect, further comprising a support member having a support part having higher rigidity than the bottom member and a holding part that holds the support part below the bottom member.

[0019] The culture vessel of the sixth aspect is provided with a support member, and a highly rigid support part is disposed below the bottom member, thereby reducing the deflection of the bottom member.

[0020] A culture vessel of a seventh aspect is the culture vessel of the sixth aspect, wherein the holding part is fitted into the main body part to hold the support part.

[0021] In the culture vessel of the seventh aspect, the holder of the support member is fitted into the main body, so that the support member can be held below the bottom member without relying on other members.

[0022] An eighth aspect of the culture vessel is the culture vessel of the sixth aspect, wherein the holding part is a base that supports the support part from below.

[0023] In the culture vessel of the eighth aspect, the holder of the support member is a base that supports the support member from below. Therefore, compared to when the holder of the support member is fitted into the main body, the support member can be held below the bottom member with a simpler configuration.

[0024] A culture vessel of a ninth aspect is the culture vessel of the first aspect, wherein the bottom member is a porous body.

[0025] In the ninth embodiment of the culture vessel, the bottom member is porous. Therefore, by immersing the culture vessel in the culture medium, the bottom member is impregnated with the culture medium, and the bottom member is immersed in the culture medium. Therefore, the culture medium is supplied to the bottom member from above and below. This improves the cell culture efficiency. In contrast, if the bottom member is not porous, the culture medium is supplied only from one side of the bottom member, resulting in relatively poor culture efficiency.

[0026] A culture vessel of a tenth aspect is the culture vessel of the first aspect, wherein the bottom member is formed using a fibrous material.

[0027] In the tenth embodiment of the culture vessel, the bottom member is formed using a fibrous material. Therefore, by immersing the culture vessel in the culture medium, the bottom member is impregnated with the culture medium, and the bottom member is immersed in the culture medium. Therefore, the culture medium is supplied to the bottom member from above and below. This improves the cell culture efficiency. In contrast, if the bottom member is not porous, the culture medium is supplied only from one side of the bottom member, resulting in relatively poor culture efficiency.

[0028] The culture vessel of the eleventh aspect is a culture vessel of the ninth or tenth aspect, which is provided with a support member having a support portion that is more rigid than the bottom member and a holding portion that holds the support portion below the bottom member, and the support portion is formed of a porous material.

[0029] In the culture vessel of the eleventh aspect, the support portion of the support member is formed of a porous material. Therefore, when the culture medium is supplied from the underside of the bottom member, the support portion is impregnated with the culture medium. Therefore, the supply of the culture medium to the bottom member is less likely to be hindered by the support portion.

[0030] A culture vessel of a twelfth aspect is the culture vessel of the first aspect, wherein the bottom member is made of a material that is biodegradable.

[0031] In the culture vessel of the twelfth aspect, the bottom member is biodegradable, so that even if cells are harvested together with the bottom member using an access device such as a cell harvesting device and then transplanted together with the bottom member into a living body, the bottom member decomposes in vivo and only the cells are transplanted.

[0032] The culture vessel of the thirteenth aspect is a culture vessel of the first aspect, in which the outer surface of an access device inserted from the opening at the upper end portion of the culture surface contacts at least a portion of the inner surface of the main body portion, thereby guiding the tip of the access device to a predetermined position on the culture surface.

[0033] In the culture vessel of the thirteenth aspect, cells can be cultured on the bottom member at the lower end of the main body. An access instrument can then be brought close to the culture surface to access the cultured cells. For example, when a cell collection instrument is used as the access instrument, cells can be collected by inserting the cell collection instrument from the upper end of the cylindrical main body. In this case, the outer surface of the access instrument contacts at least a portion of the inner surface of the culture vessel, and the tip of the access instrument is guided to a predetermined position on the culture surface.

[0034] In other words, the access range of the access instrument relative to the culture surface is positioned. When a cell acquisition instrument is used as the access instrument, the cell collection range is positioned. This makes it easier to perform cell collection work compared to a configuration in which the tip of the access instrument is not guided to a predetermined position on the culture surface, reducing the individuality and errors of the work. In addition, the cell collection range relative to the culture surface is relatively increased. As a result, the cell yield can be increased.

[0035] On the other hand, if the tip of the access device is not guided to the predetermined position on the culture surface and the access range is not positioned properly, for example, the cell collection position is not determined, which can cause deviations during cell collection and affect the health of the cell tissue. This makes the cell collection process difficult. Furthermore, since the range of movement of the tip of the cell collection device is secured on the culture surface, the cell collection range relative to the culture surface is relatively narrow, resulting in a low cell yield.

[0036] A fourteenth aspect of the culture vessel is the culture vessel of the thirteenth aspect, wherein at least a portion of the culture surface is offset outward at equal intervals from the tip of the access device.

[0037] In a fourteenth aspect of the culture vessel, at least a portion of the culture surface is offset outward at equal intervals from the tip of the access device. For example, if the tip of the access device is oval, the culture surface has a portion offset outward at equal intervals along the oval shape. This can increase the yield of cells.

[0038] On the other hand, if the culture surface is not offset outward at equal intervals from the tip of the access device, for example, if the tip of the access device is oval and the culture surface is circular, the area from which cells cannot be collected will be large along the short side of the oval, making it difficult to increase the cell yield.

[0039] A fifteenth aspect of the culture vessel is a culture vessel of the thirteenth aspect, in which, when the outer surface is in contact with the inner surface, the minimum distance between the edge of the culture surface and the tip of the access device is greater than 0.2% and less than 20% of the maximum dimension between the edges.

[0040] In the fifteenth aspect of the culture vessel, the minimum distance between the edge of the culture surface and the tip of the access device is greater than 0.2% of the maximum dimension between the edges, which provides a larger clearance around the tip of the access device than when the minimum distance is 0.2% or less, making it easier to absorb manufacturing precision errors in the culture vessel and cell collection device.

[0041] Furthermore, with this culture vessel, the minimum distance between the edge of the culture surface and the tip of the access device is less than 20% of the maximum dimension between the edges, which reduces the clearance around the tip of the access device compared to when the distance is 20% or more, thereby increasing the cell yield.

[0042] A culture vessel of the sixteenth aspect is a culture vessel of the thirteenth aspect, in which the inner surface has a plurality of contact portions formed thereon with which the outer surface can come into contact, and each contact portion guides the tip to a different position on the culture surface.

[0043] According to the culture vessel of the sixteenth aspect, a plurality of cell fragments can be collected from one culture surface.

[0044] A culture vessel of a seventeenth aspect is the culture vessel of the thirteenth aspect, wherein the shape of the culture surface is non-circular.

[0045] According to the culture vessel of the seventeenth aspect, non-circular cell fragments can be easily collected.

[0046] In an eighteenth aspect of the culture vessel, in the thirteenth aspect, the cross-sectional area of ​​the portion surrounded by the inner circumferential surface gradually decreases from the opening at the upper end toward the culture surface.

[0047] According to the culture vessel of the eighteenth aspect, an access instrument inserted from the opening at the upper end portion can be easily guided to the culture surface.

[0048] A nineteenth aspect of the method for manufacturing a culture vessel comprises the steps of: placing a bottom member, on the lower end surface of a cylindrical main body having openings at the upper and lower ends, with the opening covered; the bottom member having a culture surface made of a degradable material on which cells can be cultured; and heating the bottom member with a heat source placed between the bottom member and a buffer member, thereby heat-welding the bottom member to the lower end surface; the buffer member having non-adhesive properties that prevent the bottom member from adhering to the bottom member at the melting temperature of the bottom member.

[0049] In the nineteenth aspect of the method for producing a culture vessel, the bottom member is fixed to the lower end surface of the main body. Therefore, by placing the culture vessel on a pedestal or the like, inserting a cell collection instrument from the upper end of the cylindrical main body, and pressing the tip of the cell collection instrument against the bottom member, a reaction force is obtained from the pedestal, allowing the cultured cells to be punched out together with the bottom member and collected.

[0050] Thus, compared to a configuration in which a cell culture membrane is sandwiched and fixed between multiple components from above and below, this culture vessel eliminates the need to disassemble components when harvesting cells. Furthermore, the number of components can be reduced, reducing the risk of contamination. Furthermore, the bottom component is made of a degradable material. Therefore, after cells are harvested together with the bottom component using a cell harvesting instrument, the bottom component can be disassembled and the cells alone can be used (e.g., for biotransplantation).

[0051] Furthermore, in this method for manufacturing a culture vessel, the bottom member is heat-welded to the main body. This allows the selection of a material suited to the performance required of the bottom member. Furthermore, there is no need to use materials such as adhesives. Furthermore, in this method for manufacturing a culture vessel, the bottom member is heated by a heat source disposed with a buffer member sandwiched between the bottom member and the heat source, and the bottom member is heat-welded to the lower end surface of the main body. The buffer member is non-adhesive at the melting temperature of the bottom member, preventing the bottom member from adhering to the heat source. This prevents the molten bottom member from adhering to the heat source.

[0052] A twentieth aspect of the method for producing a culture vessel is the method for producing a culture vessel of the thirteenth aspect, wherein the buffer member contains a fluororesin.

[0053] In the method for producing a culture vessel of the twentieth aspect, the buffer member is configured to contain a fluororesin, thereby making it possible to obtain the desired effect using a general-purpose material.

[0054] According to the present disclosure, the effort required for collecting cells can be reduced.

[0055] 3A . FIG. 3B is a perspective view showing a culture mechanism according to an embodiment of the present disclosure. FIG. 3C is a cross-sectional elevation view showing a well plate and a culture vessel according to an embodiment of the present disclosure. FIG. 3D is a cross-sectional view showing a state in which cells are being cultured in a culture vessel according to an embodiment of the present disclosure. FIG. 3E is a cross-sectional view showing a state in which an access device is being inserted into a culture vessel according to an embodiment of the present disclosure. FIG. 3F is a cross-sectional view showing a state in which an access device is being inserted into a culture vessel according to an embodiment of the present disclosure. FIG. 3G is a cross-sectional view showing a state in which an access device is being inserted into a culture vessel according to an embodiment of the present disclosure. FIG. 3H is a cross-sectional view showing ... 11A , a cross-sectional view taken along line B-B in Fig. 11A; a cross-sectional view taken along line CC in Fig. 11A; a cross-sectional view taken along line B-B in Fig. 11A when the inner peripheral wall of the culture vessel according to an embodiment of the present disclosure is formed into a circular shape; and a cross-sectional view taken along line CC in Fig. 11A when the inner peripheral wall of the culture vessel according to an embodiment of the present disclosure is formed into a circular shape.

[0056] Hereinafter, a culture mechanism, a culture vessel, and a method for manufacturing a culture vessel according to embodiments of the present disclosure will be described with reference to the drawings. Components indicated by the same reference numerals in the drawings are the same components. However, unless otherwise specified in the specification, each component is not limited to one, and multiple components may be present.

[0057] Furthermore, descriptions of overlapping configurations and symbols in each drawing may be omitted. Note that the present disclosure is not limited to the following embodiments, and may be implemented by making appropriate modifications, such as omitting configurations, replacing them with different configurations, or combining one embodiment with various modified examples, within the scope of the purpose of the present disclosure.

[0058] <Culture Mechanism> The culture mechanism 10 according to the embodiment of the present disclosure shown in FIG. 1A includes an access device 20, a well plate 30, and a culture container 50.

[0059] The well plate 30 has a plurality of recesses 32 formed therein, which are holes with bottoms, and each recess 32 opens to the top surface of the well plate 30 .

[0060] 1B, the culture vessel 50 is inserted into the recess 32 from above. The culture vessel 50 has a flange 52 that protrudes laterally from the open end at the top. This flange 52 is placed on the open end of the recess 32. This allows the culture vessel 50 to be held inserted in the recess 32.

[0061] As will be described in detail later, the culture vessel 50 has a bottom member 60 at its lower end. The bottom member 60 is disposed at a distance from the bottom surface of the recess 32 when the culture vessel 50 is inserted into the recess 32.

[0062] 1C , culture solution A is stored in the recess 32, and the bottom member 60 is immersed in the culture solution A, thereby allowing, for example, cells B to be cultured on the bottom member 60. That is, the portion of the bottom member 60 that forms the bottom surface of the culture vessel 50 serves as a culture surface for cells B.

[0063] 2A, the culture vessel 50 has a cylindrical main body 50A. The material for forming the main body 50A can be selected as appropriate, but in this embodiment, MABS (transparent ABS: Methyl Methacrylate Acrylonitrile Butadiene Styrene) is used from the viewpoint of thermal welding properties, which will be described later.

[0064] The main body 50A has an opening at its upper end, at which end is formed the flange 52. The main body 50A also has an opening at its lower end, at which end is provided a bottom member 60.

[0065] As shown in FIG. 2B, the access device 20 can be inserted into the main body portion 50A through an opening formed at the top end of the main body portion 50A.

[0066] 3A, the cross-sectional area of ​​the portion surrounded by the inner peripheral surface of the wall portion 54 that forms the cylindrical shape of the main body portion 50A gradually decreases from the opening at the upper end of the main body portion 50A toward the bottom member 60. In other words, the wall portion 54 is formed to slope inward from the upper end toward the lower end of the main body portion 50A. This forms the main body portion 50A in a mortar-like shape.

[0067] Wall 54 includes upper wall 54A and lower wall 54B. Upper wall 54A has a larger inclination angle relative to the vertical direction than lower wall 54B. Therefore, the tip of access device 20 is guided to a predetermined position on base member 60 depending on the depth of insertion of access device 20 into main body 50A.

[0068] 3B , in a plan view, the wall portion 54 has a cylindrical shape formed by two identical ellipses stacked on top of each other. Specifically, the wall portion 54 has a cylindrical shape formed by arranging two ellipses side by side in the short direction, omitting the opposing longitudinal sides, and connecting the arc portions included in the ellipses.

[0069] In other words, the inner peripheral surface of the wall portion 54 has two elliptical portions C1. The two portions C1 are connected by a portion C2 that does not follow the elliptical shape. In FIG. 3B, the boundary between the portions C1 and C2 is indicated by a dashed line.

[0070] As a result, the inner peripheral surface of the wall portion 54 has a shape that allows an ellipse to be inscribed at two portions C1.

[0071] (Access Instrument) The access instrument 20 is an instrument that is inserted into the inside of the main body 50A through an opening formed at the upper end of the main body 50A and brought into close proximity with the upper surface of the bottom member 60. Various instruments such as a cell acquisition instrument, a pump, a sensor, etc. will be used as the access instrument 20. This specification will mainly describe the case where the access instrument 20 is used as a cell acquisition instrument.

[0072] As shown in Fig. 3A, the access device 20 has a cylindrical portion 22 formed at its distal end with a uniform cross section along the longitudinal direction. The distal end of the cylindrical portion 22 has a sharpened portion 22A whose outer circumferential surface is inclined toward the inner circumferential surface and whose cross-sectional area gradually decreases. The sharpened portion 22A is pressed against the base member 60, thereby punching out and collecting the base member 60 and the cells B (see Fig. 1C) cultured on the base member 60.

[0073] The outer peripheral surface of tubular portion 22 of access device 20 is partially similar in shape to the inner peripheral surface of wall portion 54 of culture vessel 50. Specifically, the outer peripheral surface of tubular portion 22 is oval. On the other hand, on the inner peripheral surface of wall portion 54, portion C1 along the oval is similar to the oval that forms the outer peripheral surface of tubular portion 22.

[0074] 3A, as shown in FIG. 3B, a portion of the outer peripheral surface of tubular portion 22 contacts portion C1 of the inner peripheral surface of wall portion 54. In other words, at point P, the outer peripheral surface of tubular portion 22 of access device 20 contacts a portion of the inner peripheral surface of main body portion 50A (i.e., portion C1).

[0075] 3C, the tip of access device 20, i.e., the tip of sharpened portion 22A, can be guided to a predetermined position on culture surface 60A of base member 60 (in other words, a determined position on culture surface 60A). Point P is the base end of sharpened portion 22A.

[0076] Here, "guiding" means regulating the posture and position of access device 20 in response to the insertion of access device 20 into main body portion 50A, and guiding the tip of sharp portion 22A to a predetermined position on culture surface 60A.

[0077] For example, in this embodiment, as access device 20 is inserted into main body 50A, the position of access device 20 is restricted so that the tip of sharpened portion 22A is parallel to culture surface 60A. Furthermore, a portion of the outer circumferential surface of tubular portion 22 of access device 20 contacts portion C1 of the inner circumferential surface of wall 54, restricting the position. The tip of sharpened portion 22A is then guided to a predetermined position on culture surface 60A, as indicated by dashed line N1 in FIG. 3C .

[0078] That is, regardless of the skill of the operator, the attitude of the access device 20 and the tip of the sharpened portion 22A are uniquely positioned.

[0079] Here, the "culture surface 60A" refers to the inner portion of the wall portion 54 of the bottom member 60. The culture surface 60A has a non-circular shape that combines portions that follow an oval shape and portions that do not. In other words, the portion surrounded by the inner peripheral surface of the wall portion 54 has a non-circular shape.

[0080] At least a portion of culture surface 60A is offset outward at an equal distance from the tip (shown by dashed line N1) of access device 20. In other words, the tip of access device 20 is positioned along a portion of the inner circumferential surface of wall 54 that is offset inward at an equal distance from portion C1.

[0081] Furthermore, when the outer surface of tubular portion 22 is in contact with the inner surface of wall portion 54 at point P shown in Figure 3A, the minimum distance W1 between the edge of culture surface 60A and the tip of access device 20 shown in Figure 3C is greater than 0.2% and less than 20% of the maximum dimension W2 between the edges of culture surface 60A.

[0082] As a specific example, the minimum distance W1 is about 0.1 mm, the maximum dimension W2 is about 2 mm, and the minimum distance W1 is about 5% of the maximum dimension W2.

[0083] Furthermore, a plurality of contact portions are formed on the inner peripheral surface of the wall portion 54, with which the outer peripheral surface of the cylindrical portion 22 can come into contact. Specifically, two portions C1 shown in FIG. 3B are contact portions with which the outer peripheral surface of the cylindrical portion 22 can come into contact.

[0084] Each portion C1 guides the tip of access device 20 to a different position on culture surface 60A. Specifically, portion C1 on the right side of Fig. 3B guides the tip of access device 20 to the portion indicated by dashed line N1 in Fig. 3C. Meanwhile, portion C1 on the left side of Fig. 3B guides the tip of access device 20 to the portion indicated by dashed line N2 in Fig. 3C.

[0085] (Bottom Member) The bottom member 60 is made of a degradable material. A "degradable material" is, for example, a material that undergoes hydrolysis. Examples of hydrolyzable materials include materials that undergo hydrolysis in vivo (hereinafter referred to as "biodegradation"). Preferably, the bottom member 60 is made of a biocompatible material. A "biocompatible material" is a material that is unlikely to cause side effects on the living body in vivo, regardless of whether it biodegrades or not.

[0086] In this embodiment, an example will be described in which PLGA (Polylactide-co-glycolide), a biodegradable material, is used as the base member 60. PLGA with a DL-lactide / glycolide ratio of approximately 1:1 can be used. From the viewpoint of fusion, the surface free energy of the MABS forming the main body portion 50A is −10 mJ / m 2 or more, and +10mJ / m 2 From the viewpoint of improving the fusion property, the surface of the MABS may be plasma-treated or may be provided with fine irregularities.

[0087] The base member 60 is formed as a porous body having a thickness of 20 to 50 μm by layering membranes made of fibrous (so-called nanofiber) PLGA having a diameter of approximately 150 to approximately 650 nm and knitted to an average mesh size of 1 μm.

[0088] 3A, the bottom member 60 is bonded to the lower end surface of the wall portion 54 of the main body portion 50A. There are no particular limitations on the method for bonding the bottom member 60 to the main body portion 50A, but in this embodiment, heat welding is used. The heat welding method is also not particularly limited, and examples include a method using a heat block, or a method using ultrasound or a laser, but in this embodiment, a heat block is used.

[0089] (Heat Welding Method) To heat weld the bottom member 60 to the main body portion 50A, for example, a mold 70 as a heat block and a fluororesin sheet 74 as a buffer member are used, as shown in FIG.

[0090] Specifically, first, the wall portion 54 of the main body portion 50A is positioned upside down relative to the usage posture (in other words, positioned so that the lower end surface of the wall portion 54 in the posture during cell culture becomes the upper end surface).

[0091] Then, the bottom member 60 is placed on the upper end surface of the wall portion 54 in this position. The bottom member 60 is large enough to protrude outward from the outer edge of the upper end surface of the wall portion 54. The bottom member 60 is also placed so as to cover the opening of the main body portion 50A surrounded by the wall portion 54.

[0092] Then, with the fluororesin sheet 74 sandwiched between the bottom member 60 and the mold 70, the mold 70 is heated to transfer heat from the mold 70 to the bottom member 60. The bottom member 60 is then pressed against the wall portion 54. This causes the bottom member 60 to be thermally welded to the main body portion 50A. Any portion of the bottom member 60 that protrudes outside the main body portion 50A may be cut off as appropriate.

[0093] During thermal welding, the temperature of the mold 70 is 200°C, the pressing force is 20 N, and the pressing time is 1 second. These temperature, pressing force, and pressing time can be changed appropriately depending on their respective values. The thickness of the fluororesin sheet 74 is not particularly limited, but is set to 75 μm, for example. This thickness can be changed appropriately depending on the heating conditions and pressing conditions.

[0094] The fluororesin sheet 74 preferably covers the entire surface of the bottom member 60. This can suppress the transfer of radiant heat from the mold 70 to the bottom member 60. Note that materials other than fluororesin may be used as the buffer member. However, it is preferable to use a material that is non-adhesive at the melting temperature of the bottom member 60 so that the bottom member 60 does not adhere to it.

[0095] 5 and 6 show the positional relationship between the mold 70 and the wall 54 of the main body 50A when the mold 70 presses the main body 50A. Note that these figures omit the illustration of the bottom member 60 and the fluororesin sheet 74. Note that Fig. 6 shows an enlarged view of the area indicated by 6A in Fig. 5 and a cross-sectional view of the portion indicated by line 6B-6B in Fig. 5.

[0096] As shown in FIG. 5, in the mold 70, the heating portion 72 facing the wall portion 54 does not face the inner portion of the end face of the wall portion 54, but faces the outer portion.

[0097] Specifically, the thickness W3 of the wall portion 54 shown in Figure 6 is, for example, 1.0 mm, and the thickness W4 of the heating portion 72 is, for example, 0.9 mm. The heating portion 72 is disposed offset outward by a width W5 relative to the wall portion 54. This width W5 is, for example, 0.6 mm. As a result, the overlap width W6 between the wall portion 54 and the heating portion 72 is 0.4 mm.

[0098] A chamfer R is formed between the lower end surface of the heating portion 72 and the inner peripheral wall, and the radius of the chamfer is 0.3 mm. As a result, the thickness W7 of the portion of the overlap width W6 between the wall portion 54 and the heating portion 72 where the wall portion 54 and the lower end surface of the heating portion 72 face each other in parallel is 0.1 mm.

[0099] (Supporting Member) A supporting member 80 shown in Fig. 7 can be attached to the culture vessel 50. The supporting member 80 includes a supporting portion 80A and a holding portion 80B.

[0100] The support portion 80A is made of a porous material with a pore diameter of approximately 0.4 μm, and is a membrane material with higher rigidity than the bottom member 60. The material from which the support portion 80A is made is not particularly limited, but polycarbonate, for example, can be used. The support portion 80A is heat-welded to the lower end surface of the holding portion 80B.

[0101] The phrase "higher rigidity than the bottom member 60" means that the bottom member 60 is less likely to bend in the out-of-plane direction (downward in the culture posture of the cells) than the bottom member 60.

[0102] The holding portion 80B is a cylindrical member that holds the support portion 80A below the bottom member 60. The holding portion 80B has an opening at its upper end, and a flange 82 that protrudes laterally is provided at the edge of the upper opening. The holding portion 80B also has an opening at its lower end, and the support portion 80A is provided at this opening edge.

[0103] 8A, the culture vessel 50 can be inserted into the opening formed at the upper end of the holder 80B. The cross-sectional area of ​​the portion surrounded by the inner circumferential surface of the wall 84 that forms the cylindrical shape of the holder 80B gradually decreases from the opening at the upper end of the holder 80B toward the support part 80A. In other words, the wall 84 is formed to slope inward from the upper end toward the lower end of the holder 80B. This forms the holder 80B in a mortar shape.

[0104] The wall portion 84 includes an upper wall portion 84A and a lower wall portion 84B. The upper wall portion 84A has a larger inclination angle with respect to the vertical direction than the lower wall portion 84B.

[0105] As shown in Figure 8B, when the culture vessel 50 is inserted into the holding portion 80B of the support member 80, the upper wall portion 54A of the main body portion 50A of the culture vessel 50 is positioned along and in contact with the upper wall portion 84A of the holding portion 80B of the support member 80.

[0106] In this state, the bottom member 60 of the culture vessel 50 is disposed adjacent to or in contact with the support portion 80A of the support member 80. In this manner, the holding portion 80B is fitted into the main body portion 50A to hold the support portion 80A below the bottom member 60.

[0107] Note that, regardless of whether the support member 80 is used or not, ribs 54C may be provided along the vertical direction on the wall 54 of the culture vessel 50. Fig. 2A shows an example in which the ribs 54C are not provided, and Fig. 7 shows an example in which the ribs 54C are provided.

[0108] Providing the ribs 54C increases the torsional rigidity of the wall portion 54, thereby suppressing deformation of the bottom member 60 joined to the wall portion 54. Furthermore, providing the ribs 54C brings the ribs 54C into contact with the upper wall portion 84A of the holder 80B. This reduces the contact area between the culture vessel 50 and the support member 80 compared to when the ribs 54C are not provided, and compressive stress acts locally between the culture vessel 50 and the support member 80. As a result, the ribs 54C bite into the support member 80, fixing them to each other.

[0109] (Modification of Support Member) The culture vessel 50 can also be supported by a support member 90 shown in Fig. 9. The support member 90 includes a support portion 90A and a holding portion 90B.

[0110] The support portion 90A is made of a porous material with a pore diameter of approximately 0.4 μm, and is a membrane material with higher rigidity than the bottom member 60. The material from which the support portion 90A is made is not particularly limited, but polycarbonate, for example, can be used. The support portion 90A is heat-welded to the upper end surface of the holding portion 90B.

[0111] The holding portion 90B is a base that holds the support portion 90A below the bottom member 60. The holding portion 90B has three legs 92 and an annular member 94 supported by the legs 92. The support portion 90A is heat-welded to the upper end surface of the annular member 94 and is supported from below by the holding portion 90B.

[0112] When using this support member 90, as shown in Figure 10, the support member 90 is placed in the recess 32 of the well plate 30, and the culture vessel 50 is placed above it. It is preferable to use a support member 90 having a higher specific gravity than the culture solution A (see Figure 1C). Note that the holding part 90B may be configured to hold the support part 90A using only the legs 92, omitting the annular member 94.

[0113] <Actions and Effects> In the culture vessel 50 according to the embodiment of the present disclosure, as shown in Fig. 1C, cells B can be cultured on the bottom member 60 at the lower end of the main body 50A. Then, as shown in Fig. 3A, an access tool 20 can be brought close to the culture surface of the cultured cells B (cells B are not shown in figures other than Fig. 1C).

[0114] For example, when a cell collection device is used as access device 20, cells B can be collected by inserting the cell collection device from the upper end of cylindrical main body 50A. At this time, the outer surface of cylindrical portion 22 of access device 20 (the outer surface indicated by point P in FIG. 3A) comes into contact with at least a portion of the inner surface of culture vessel 50 (portion C1 in FIG. 3B). Then, the tip of access device 20, indicated by dashed line N1 in FIG. 3C, is guided to a predetermined position on culture surface 60A.

[0115] That is, the access range of the access device 20 relative to the culture surface 60A is positioned. When a cell acquisition device is used as the access device 20, the cell collection range is positioned. This allows the cell collection work to be easily performed, reducing the individuality of the work and errors. Furthermore, the cell collection range relative to the culture surface 60A is increased. As a result, the cell yield can be increased.

[0116] On the other hand, if the tip of the access device is not guided to the predetermined position on the culture surface and the access range is not positioned properly, for example, the cell collection position is not determined, which can cause deviations during cell collection and affect the health of the cell tissue. This makes the cell collection process difficult. Furthermore, since the range of movement of the tip of the cell collection device is secured on the culture surface, the cell collection range relative to the culture surface is relatively narrow, resulting in a low cell yield.

[0117] 3C , in the culture vessel 50 according to the embodiment of the present disclosure, at least a portion of the culture surface 60A is offset outward at an equal interval from the tip of the access device 20. In other words, at least a portion of the inner circumferential surface of the wall portion 54 of the culture vessel 50 is disposed at a position offset outward at an equal interval from the tip of the access device 20.

[0118] For example, in this embodiment, the tip of access device 20 has an oval shape, and culture surface 60A has portions offset outward at equal intervals along the oval shape, thereby increasing the yield of cells.

[0119] In contrast, as shown in the "Comparative Example" in Figure 3D, if culture surface 600A is not offset outward at equal intervals from tip N1 of access device 20, for example, if the tip of access device 20 is oval and culture surface 600A (the inner peripheral surface of wall portion 540) is circular, a large area (e.g., area L1) will be formed along the short side of the oval from which cells cannot be collected. This makes it difficult to increase the cell yield.

[0120] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the minimum distance W1 between the edge of the culture surface 60A and the tip of the access device is greater than 0.2% of the maximum dimension W2 between the edges. This provides a wider clearance around the tip of the cell collection device compared to when the minimum distance W1 is 0.2% or less, making it easier to absorb manufacturing precision errors in the culture vessel 50 and the access device 20.

[0121] Furthermore, with this culture vessel 50, the minimum distance W1 between the edge of the culture surface and the tip of the access device is less than 20% of the maximum dimension W2 between the edges, which reduces the clearance around the tip of the access device 20 compared to when the distance is 20% or more, thereby increasing the cell yield.

[0122] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the inner circumferential surface of the wall portion 54 is formed with a plurality of contact portions (portions C1) that can come into contact with the outer circumferential surface of the tubular portion 22. Each portion C1 guides the tip of the access device 20 to a different position on the culture surface indicated by the dashed dotted lines N1 and N2 in Figure 3C. This allows multiple cell fragments to be collected from one culture surface 60A.

[0123] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the culture surface 60A has a non-circular shape that is a combination of oval portions, which makes it easier to collect oval cell fragments.

[0124] 3A , in culture vessel 50 according to an embodiment of the present disclosure, the cross-sectional area of ​​the portion surrounded by the inner circumferential surface of wall portion 54 that forms main body portion 50A into a cylindrical shape gradually decreases from the opening at the upper end of main body portion 50A toward bottom member 60. This makes it easier to guide access device 20 inserted from the opening at the upper end of main body portion 50A to culture surface 60A.

[0125] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is formed of a degradable material, so that after cells B (see FIG. 1C ) are collected together with the bottom member 60 using an access device, the bottom member 60 can be degraded and the cells alone can be used (e.g., for biotransplantation).

[0126] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is biodegraded. As a result, even if cells B are collected together with the bottom member 60 using the access device 20 and then transplanted together with the bottom member 60 into a living body, the bottom member 60 decomposes in vivo, and only the cells are transplanted. In other words, the bottom member 60, which is not decomposed when cells B are collected and transplanted, decomposes in vivo after transplantation into a living body.

[0127] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is formed as a porous body by arranging membrane bodies made of woven fibrous PLGA in layers.

[0128] 1C, by immersing the culture vessel 50 in the culture solution A, the bottom member 60 is impregnated with the culture solution A, and the bottom member 60 is immersed in the culture solution A. As a result, the culture solution A is supplied to the bottom member 60 from above and below. This improves the culture efficiency of the cells B. In contrast, if the bottom member is not porous, the culture solution A is supplied only from one side of the bottom member, resulting in relatively poor culture efficiency.

[0129] 1B, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is fixed to the lower end of the main body portion 50A and forms the bottom of the main body portion 50A. Therefore, by placing the culture vessel 50 on a base or the like, inserting a cell collection tool, which is an example of an access tool 20, from the upper end of the cylindrical main body portion 50A, and pressing the tip of the cell collection tool against the bottom member 60, a reaction force is obtained from the base, allowing the cultured cells B (see FIG. 1C) to be punched out and collected together with the bottom member 60.

[0130] Thus, compared to a configuration in which the cell culture membrane is fixed by sandwiching it between multiple members from above and below, this culture vessel 50 can eliminate the need to disassemble the members when harvesting cells B. In other words, it reduces the effort required when harvesting cells B. Furthermore, because the number of members can be reduced, there is less risk of contamination.

[0131] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is bonded to the lower end surface of the main body portion 50A. That is, the bottom member 60 is formed separately from the main body portion 50A. This allows a material (PLGA in this embodiment) to be selected according to the performance required for the bottom member 60.

[0132] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, the bottom member 60 is heat-welded to the main body portion 50A. This eliminates the need to use materials such as adhesives. As a result, no other materials are interposed between the bottom member 60 and the main body portion 50A, reducing the number of parts.

[0133] Furthermore, in the culture vessel 50 according to the embodiment of the present disclosure, when the bottom member 60 is heat-welded, it is positioned in the positional relationship with the mold 70 shown in Fig. 6. As a result, the bottom member 60 is heat-welded to the outer portion of the lower end surface of the main body 50A from a position offset outward from the culture surface 60A. Therefore, the influence of heat on the culture surface 60A can be reduced compared to when there is no offset.

[0134] In addition, in the culture vessel 50 according to the embodiment of the present disclosure, the surface free energy of the main body portion 50A is −10 mJ / m 2 or more and +10mJ / m 2As a result, the surface free energy of the main body 50A is −10 mJ / m 2 If it is smaller than this, it will be +10mJ / m 2 Compared with a larger diameter, the bondability by thermal welding is improved.

[0135] Furthermore, the culture vessel 50 according to the embodiment of the present disclosure can be used in combination with the support member 80 shown in Fig. 7 or the support member 90 shown in Fig. 9. This can reduce bending of the bottom member 60.

[0136] Of these, the support member 80 has the holding portion 80B fitted into the main body portion 50A of the culture vessel 50. Therefore, the support portion 80A can be held below the bottom member 60 without relying on other members.

[0137] On the other hand, the support member 90 has a holding portion 90B which is a base that supports the support portion 90A from below. Therefore, compared to when the holding portion of the support member is fitted into the main body portion, the support portion 90A can be held below the bottom member 60 with a simpler configuration.

[0138] Furthermore, the support portion 80A of the support member 80 and the support portion 90A of the support member 90 are formed of a porous material. Therefore, as shown in Fig. 1C, when culture solution A is supplied from the underside of the bottom member 60, the support portions 80A and 90A are impregnated with the culture solution A. Therefore, the supply of culture solution to the bottom member 60 is less likely to be hindered by the support portions 80A and 90A.

[0139] Furthermore, in the manufacturing method of the culture vessel 50 according to an embodiment of the present disclosure (the method of thermally welding the bottom member 60 to the main body portion 50A), as shown in FIG. 4 , the bottom member 60 is heated by a heat source (mold 70) arranged with a fluororesin sheet 74 sandwiched between the bottom member 60 and the mold 70 as a buffer member, and the bottom member 60 is thermally welded to the lower end surface of the main body portion 50A.

[0140] The fluororesin sheet 74 has non-adhesive properties that prevent the bottom member 60 from adhering to it at the melting temperature of the bottom member 60. Therefore, it is possible to prevent the molten bottom member 60 from adhering to the mold 70.

[0141] Furthermore, in the manufacturing method of the culture vessel 50 according to the embodiment of the present disclosure, the fluororesin sheet 74 is used as a buffer member, which has the effect of preventing the molten bottom member 60 from adhering to the mold 70 using a general-purpose material.

[0142] <Other Embodiments> In the above embodiment, at least a portion of culture surface 60A of culture vessel 50 is offset outward at equal intervals from the tip of access instrument 20, but the embodiments of the present disclosure are not limited to this.

[0143] For example, even if the outer peripheral surface of access device 20 contacts main body 50A of culture vessel 50 and the tip of access device 20 is guided to a predetermined position on culture surface 60A, it is not necessary to offset at "equidistant intervals." In other words, the shape of the tip of access device 20 and the shape of culture surface 60A can be selected appropriately depending on the shape of the desired cell fragment.

[0144] Furthermore, in the above embodiment, a portion of the outer peripheral surface of tubular portion 22 of access device 20 contacts portion C1 of the inner peripheral surface of wall portion 54, and the posture of access device 20 and the tip of sharp portion 22A are “uniquely” positioned, but embodiments of the present disclosure are not limited to this.

[0145] For example, when a portion of the outer peripheral surface of the tubular portion 22 is in contact with portion C1 of the inner peripheral surface of the wall portion 54, the tip of the sharp portion 22A may be guided to a "position offset" on the culture surface 60A from the position indicated by the dotted line N1 in Figure 3C.

[0146] That is, when the tip of sharpened portion 22A is in contact with culture surface 60A, a range of motion (play) may be ensured between access device 20 and main body 50A. Even in this embodiment, the range of motion of access device 20 is restricted to the amount of motion within the offset range described above. Therefore, cell fragments of the desired size can be easily collected even if hand shake or the like occurs.

[0147] Furthermore, as described above, the method for guiding the tip of the access device 20 to a predetermined position on the culture surface 60A is to bring a portion of the outer surface of the tubular portion 22 of the access device 20 into contact with the portion C1 of the inner surface of the wall portion 54.

[0148] In this embodiment, the outer surface of the cylindrical portion 22 of the access device 20 (the outer surface indicated by point P in FIG. 3A ) is in “line contact” with the portion C1 of the culture vessel 50, but the embodiments of the present disclosure are not limited to this.

[0149] For example, a method of fixing any three points on the access device 20 to the inner periphery of the main body portion 50A (point contact) or a method of fixing any surface on the access device 20 to the inner periphery of the main body portion 50A (surface contact) may be adopted.

[0150] Furthermore, in the above embodiment, contact portions with which the outer surface of the access device 20 can come into contact are formed in "multiple locations" on the inner surface of the main body 50A of the culture vessel 50, but the embodiments of the present disclosure are not limited to this.

[0151] For example, as in the culture vessel 51 shown in FIGS. 11A to 11C, only one contact portion that can come into contact with the outer circumferential surface of the access device 20 may be formed on the inner circumferential surface of the wall portion 51A.

[0152] In this embodiment, the inner peripheral surface of wall 51A of culture vessel 51 also has an oval-shaped inner peripheral surface relative to the outer peripheral surface of oval-shaped access device 20. At point P shown in Fig. 11A, as shown in Fig. 11B, the inner peripheral surface of wall 51A contacts the outer peripheral surface of access device 20. Furthermore, as shown in Fig. 11C, culture surface 60A as a whole is offset outward at equal intervals from the tip of access device 20.

[0153] In the above embodiment, the inner circumferential surface (culture surface 60A) of wall portion 54 of culture vessel 50 and the outer circumferential surface of tubular portion 22 of access device 20 are non-circular, but the embodiments of the present disclosure are not limited to this. For example, they may be circular, as in the inner circumferential surface of culture vessel 53 and the outer circumferential surface of access device 21 shown in Figures 11D and 11E.

[0154] In the above embodiment, the cross-sectional area of ​​the portion of the culture vessel 50 surrounded by the inner circumferential surface of the wall portion 54 gradually decreases from the opening at the upper end of the main body portion 50A toward the culture surface 60A, but the embodiment of the present disclosure is not limited to this. For example, the wall portion 54 may be cylindrical in shape, with the cross-sectional area of ​​the portion surrounded by the inner circumferential surface being constant.

[0155] Furthermore, in the above embodiment, the outer peripheral surface of access device 20 contacts at least a portion of the inner peripheral surface of main body portion 50A, and the tip of access device 20 of culture vessel 50 is guided to a predetermined position on culture surface 60A. However, the embodiment of the present disclosure is not limited to this, and the outer peripheral surface of access device 20 does not have to contact the inner peripheral surface of main body portion 50A.

[0156] In the above embodiment, the bottom member 60 is porous, but the present disclosure is not limited to this. For example, if culture solution A shown in FIG. 1C is poured into the culture vessel 50 from above the bottom member 60, or if the entire culture vessel 50 is immersed in culture solution A, cells B can be cultured without using a porous bottom member 60.

[0157] In other words, the bottom member 60 does not need to be made of a membrane body made of woven fibrous PLGA, but may be made of a membrane material with no openings, or may not be made of PLGA, and an appropriate material can be selected depending on the purpose.

[0158] In the above embodiment, the bottom member 60 is heat-welded to the lower end surface of the main body 50A at a position offset outward from the culture surface 60A, but the embodiment of the present disclosure is not limited to this. For example, the bottom member 60 may be heat-welded to the entire lower end surface of the main body 50A. The same applies when an adhesion method other than heat welding is used.

[0159] In the above embodiment, the bottom member 60 is bonded to the lower end surface of the main body 50A, but the embodiment of the present disclosure is not limited to this. For example, if the bottom member 60 is made of a hard material similar to that of the main body 50A (e.g., polycarbonate), the bottom member 60 may be fixed by fitting to the main body 50A.

[0160] In the above embodiment, the surface free energy of the main body 50A is set to −10 mJ / m 2 or more and +10mJ / m 2 Although the following description is given, the embodiment of the present disclosure is not limited to this. If the bottom member 60 is not heat-welded to the lower end surface of the main body portion 50A, the surface free energy can be freely set.

[0161] Furthermore, in the above embodiment, the bottom member 60 is formed of a degradable material, but the embodiment of the present disclosure is not limited to this. Even if the bottom member 60 is not made of a degradable material, cells B can be cultured on the surface of the bottom member 60.

[0162] Furthermore, in the above embodiment, the culture vessel 50 is described as having a configuration including the bottom member 60, but the culture vessel of the present disclosure need only have an opening formed at the lower end of the main body portion 50A that can be covered by the bottom member 60, and does not necessarily need to include the bottom member 60. In this way, the present disclosure can be embodied in various forms.

[0163] <Supplementary Note 1> (((1))) A culture vessel comprising: a main body portion formed in a cylindrical shape with an opening at an upper end; and a bottom member fixed to a lower end of the main body portion to form the bottom of the main body portion, the bottom member having a culture surface on which cells can be cultured, wherein the bottom member is formed from a degradable material.

[0164] (((2))) The culture vessel according to (((1))), wherein the bottom member is adhered to a lower end surface of the main body portion.

[0165] (((3))) The culture vessel according to (((2))), wherein the bottom member is heat-welded to the lower end surface.

[0166] (((4))) The culture vessel according to (((3))), wherein the bottom member is heat-welded to an outer portion of the lower end surface from a position offset outward from the culture surface.

[0167] (((5))) The surface free energy of the main body is −10 mJ / m relative to the surface free energy of the bottom member. 2 or more and +10mJ / m 2The culture vessel according to (((3))), which is:

[0168] (((6))) A culture vessel according to any one of (((1))) to (((5))), comprising a support member having a support portion having higher rigidity than the bottom member, and a holding portion that holds the support portion below the bottom member.

[0169] (((7))) The culture vessel according to (((6))), wherein the holding part is fitted into the main body part to hold the support part.

[0170] (((8))) The culture vessel according to (((6))), wherein the holding part is a base that supports the support part from below.

[0171] (((9))) The culture vessel according to any one of (((1))) to (((8))), wherein the bottom member is a porous body.

[0172] (((10))) The culture vessel according to any one of (((1))) to (((9))), wherein the bottom member is formed using a fibrous material.

[0173] (((11))) A culture vessel according to (((9))) or (((10))), comprising a support member having a support part having higher rigidity than the bottom member, and a holding part that holds the support part below the bottom member, wherein the support part is formed of a porous body.

[0174] (((12))) The culture vessel according to any one of (((1))) to (((11))), wherein the bottom member is made of a material that is biodegradable.

[0175] (((13))) A culture vessel as described in (((1))), in which the outer surface of an access instrument to the culture surface inserted from the opening at the upper end portion contacts at least a portion of the inner surface of the main body portion, thereby guiding the tip of the access instrument to a predetermined position on the culture surface.

[0176] (((14))) The culture vessel of (((13))), wherein at least a portion of the culture surface is offset outwardly at equal intervals from the tip of the access device.

[0177] (((15))) A culture vessel according to (((13))) or (((14))), wherein, with the outer peripheral surface in contact with the inner peripheral surface, the minimum distance between the edge of the culture surface and the tip of the access device is greater than 0.2% and less than 20% of the maximum dimension between the edges.

[0178] (((16))) A culture vessel described in any of (((13))) to (((15))), wherein a plurality of contact portions are formed on the inner circumferential surface with which the outer circumferential surface can come into contact, and each contact portion guides the tip to a different position on the culture surface.

[0179] (((17))) The culture vessel according to any one of (((13))) to (((16))), wherein the culture surface has a non-circular shape.

[0180] (((18))) A culture vessel according to any one of (((13))) to (((18))), wherein the cross-sectional area of ​​the portion surrounded by the inner peripheral surface gradually decreases from the opening at the upper end toward the culture surface.

[0181] (((19))) A method for manufacturing a culture vessel, comprising the steps of: placing a bottom member, having a culture surface made of a degradable material on the lower end surface of a cylindrical main body having openings at the upper and lower ends, with the openings covered; heating the bottom member with a heat source arranged with a buffer member sandwiched between the bottom member and the heat source, and heat welding the bottom member to the lower end surface; wherein the buffer member is non-adhesive so that the bottom member does not adhere to the buffer member at the melting temperature of the bottom member.

[0182] (((20))) The method for producing a culture vessel according to (((19))), wherein the buffer member is configured to contain a fluororesin.

[0183] <Supplementary Note 2> (((1))) A culture vessel having a cylindrical main body portion with an opening at an upper end, wherein the lower end of the main body portion is provided with a bottom member having a culture surface on which cells can be cultured, or an opening that can be covered with a bottom member having a culture surface on which cells can be cultured is formed, and wherein an outer surface of an access instrument inserted from the upper end opening contacts at least a part of the inner circumferential surface of the main body portion, thereby guiding the tip of the access instrument to a predetermined position on the culture surface.

[0184] (((2))) The culture vessel according to (((1))), wherein at least a portion of the culture surface is offset outward at equal intervals from the tip of the access device.

[0185] (((3))) The culture vessel described in (((1))) or (((2))), wherein, when the outer peripheral surface is in contact with the inner peripheral surface, the minimum distance between the edge of the culture surface and the tip of the access device is greater than 0.2% and less than 20% of the maximum dimension between the edges.

[0186] (((4))) A culture vessel described in any of (((1))) to (((3))), wherein the inner circumferential surface has a plurality of contact portions formed thereon with which the outer circumferential surface can come into contact, and each contact portion guides the tip to a different position on the culture surface.

[0187] (((5))) The culture vessel according to any one of (((1))) to (((4))), wherein the culture surface has a non-circular shape.

[0188] (((6))) A culture vessel according to any one of (((1))) to (((5))), wherein the cross-sectional area of ​​the portion surrounded by the inner peripheral surface gradually decreases from the opening at the upper end toward the culture surface.

[0189] (((7))) The culture vessel according to any one of (((1))) to (((6))), wherein the bottom member is formed from a degradable material.

[0190] (((8))) The culture vessel according to (((7))), wherein the bottom member is made of a material that is biodegradable.

[0191] (((9))) The culture vessel according to any one of (((1))) to (((8))), wherein the bottom member is a porous body.

[0192] (((10))) The culture vessel according to any one of (((1))) to (((9))), wherein the bottom member is formed using a fibrous material.

[0193] (((11))) A culture mechanism comprising: a culture vessel having a cylindrical main body portion with an opening at an upper end, and a bottom member disposed at a lower end of the main body portion and having a culture surface on which cells can be cultured; and an access instrument that can be inserted through the upper opening, the outer surface of which comes into contact with at least a part of the inner surface of the culture vessel, and the tip of which is guided to a predetermined position on the culture surface.

[0194] The disclosures of Japanese Patent Application No. 2023-206456, filed on December 6, 2023, and Japanese Patent Application No. 2023-207161, filed on December 7, 2023, are incorporated herein by reference in their entirety. All documents, patent applications, and technical standards mentioned herein are incorporated herein by reference to the same extent as if each individual document, patent application, and technical standard was specifically and individually indicated to be incorporated by reference.

Claims

1. A culture vessel comprising: a main body portion formed in a cylindrical shape with an opening at an upper end; and a bottom member fixed to the lower end of the main body portion to form the bottom of the main body portion, the bottom member having a culture surface on which cells can be cultured, the bottom member being formed from a degradable material.

2. The culture vessel according to claim 1, wherein the bottom member is bonded to the lower end surface of the main body portion.

3. The culture vessel according to claim 2, wherein the bottom member is heat welded to the lower end surface.

4. The culture vessel according to claim 3, wherein the bottom member is heat welded to an outer portion of the lower end surface from a position offset outward from the culture surface.

5. The surface free energy of the main body is -10 mJ / m 2 or more and +10mJ / m 2 The culture vessel according to claim 3 , wherein:

6. The culture vessel according to claim 1, comprising a support member having a support part having higher rigidity than said bottom member, and a holding part that holds said support part below said bottom member.

7. The culture vessel according to claim 6, wherein the holding part is fitted into the main body part to hold the support part.

8. The culture vessel according to claim 6, wherein the holding part is a base that supports the support part from below.

9. The culture vessel according to claim 1, wherein the bottom member is a porous body.

10. The culture vessel according to claim 1, wherein the bottom member is formed using a fibrous material.

11. A culture vessel as described in claim 9 or 10, comprising a support member having a support portion having higher rigidity than the bottom member and a holding portion that holds the support portion below the bottom member, the support portion being formed of a porous material.

12. The culture vessel according to claim 1, wherein the bottom member is made of a material that is decomposed by biodegradation.

13. A culture vessel as described in claim 1, wherein the outer surface of an access instrument to the culture surface inserted from the opening at the upper end portion contacts at least a portion of the inner surface of the main body portion, thereby enabling the tip of the access instrument to be guided to a predetermined position on the culture surface.

14. The culture vessel of claim 13, wherein at least a portion of the culture surface is offset outwardly and equidistantly from the tip of the access device.

15. The culture vessel according to claim 13, wherein, with the outer circumferential surface in contact with the inner circumferential surface, the minimum distance between the edge of the culture surface and the tip of the access device is greater than 0.2% and smaller than 20% of the maximum dimension between the edges.

16. The culture vessel according to claim 13, wherein a plurality of contact portions are formed on the inner circumferential surface with which the outer circumferential surface can come into contact, and each contact portion guides the tip to a different position on the culture surface.

17. The culture vessel according to claim 13, wherein the culture surface has a non-circular shape.

18. The culture vessel according to claim 13, wherein the cross-sectional area of ​​the portion surrounded by the inner peripheral surface gradually decreases from the opening at the upper end toward the culture surface.

19. A method for manufacturing a culture vessel, comprising the steps of: placing a bottom member, on the lower end surface of a cylindrical main body having openings at the upper and lower ends, with the openings covered; and heating the bottom member with a heat source arranged with a buffer member sandwiched between the bottom member and the heat source, thereby heat-welding the bottom member to the lower end surface, wherein the buffer member has non-adhesive properties that prevent the bottom member from adhering to the bottom member at the melting temperature of the bottom member.

20. The method for manufacturing a culture vessel according to claim 19, wherein the buffer member is composed of a fluororesin.

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