Transport container for protecting grafts and method of using the transport container

Abstract

The purpose of the present invention is to provide a transport container capable of protecting a transplant by capturing air bubbles remaining therein using a simple mechanism and a simple operation. This transport container is for protecting a transplant and comprises a lid for sealing a container for housing a transplant. The lid, in a state of being attached to the container, has a protruding section that protrudes toward the space inside the container. A generally annular groove section is provided in a lower surface of the protruding section. It is possible to obtain a liquid-tight state by pushing, with the lid, gas and liquid out from the container having housed therein a transplant and liquid, and to capture air bubbles in the liquid by the groove section in the lower surface.

Description

[Technical Field]

[0001] The present invention relates to a shipping container for protecting an implant and a method for using the shipping container. [Background technology]

[0002] In recent years, new regenerative medicines have been developed as a solution for treating severe heart failure. One example is a method in which sheet-shaped cell cultures created using temperature-responsive culture dishes using tissue engineering are applied to the heart surface in cases of severe myocardial infarction, etc. The method using sheet-shaped cell cultures makes it possible to safely transplant large amounts of cells over a wide area, and is particularly useful for treating heart diseases (e.g., heart failure, particularly chronic heart failure) accompanied by myocardial infarction (including chronic heart failure associated with myocardial infarction), dilated cardiomyopathy, ischemic cardiomyopathy, and systolic dysfunction (e.g., left ventricular systolic dysfunction).

[0003] To apply such sheet-shaped cell cultures clinically, for example, the prepared sheet-shaped cell cultures must be placed in a container together with a preservation solution and transported to an intensive care unit where the transplant will take place, etc. However, sheet-shaped cell cultures have low absolute physical strength, and vibrations that occur when transporting the container can cause wrinkles, tears, or damage, so this transport process requires advanced techniques and meticulous care.

[0004] To meet these needs, various methods and containers have been developed. For example, the storage and transportation container for membranous tissues described in Patent Document 1 below fills the storage compartment with a preservation solution to the extent that a gas layer is not formed, thereby preventing the preservation solution from rippling or flowing. As a result, vibrations are not transmitted to the membranous tissue, preventing damage to the membranous tissue. Furthermore, a clip-shaped connecting mechanism that elastically clamps the container body and the lid member has been devised to make the connection easier.

[0005] The packaging container for transporting biological samples and the like described in Patent Document 2 has a lid with a recessed portion that protrudes toward the bottom of the packaging container body, and the recessed portion has a rim that protrudes toward the bottom of the packaging container body, and the rim is provided with an air escape hole. When the lid is sealed, the air inside the recessed portion moves outward through the air escape hole, thereby reducing the space within the sample container in which the culture medium can move due to vibrations, impacts, and tilting that occur to the sample container during transportation.

[0006] The device for storing fragile objects described in Patent Document 3 includes a lid member, which has a convex portion that protrudes upward into the lower space when attached to the container, the horizontal cross-sectional area of ​​the convex portion decreasing as it extends vertically upward, and a communication mechanism that can be opened and closed is provided at the top of the convex portion, and the lid member can push out gas and liquid inside the container that stores fragile objects and liquid, thereby making the lower space liquid-tight. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Japanese Patent Application Laid-Open No. 2012-130311 [Patent Document 2] Japanese Patent Application Laid-Open No. 2013-128457 [Patent Document 3] International Publication No. 2019 / 017466 Summary of the Invention [Problem to be solved by the invention]

[0008] In devices that push out gas, air bubbles may remain inside the lid during the sealing process. It is difficult to visually check for air bubbles during the process, and if air bubbles are discovered after sealing, they are difficult to remove.

[0009] The present invention has been made in consideration of these points, and its object is to provide a transport container that has a simple mechanism and can capture air bubbles remaining in the container with simple operation, thereby protecting the graft. [Means for solving the problem]

[0010] That is, the present invention relates to the following. [1] A transport container for protecting a graft, comprising a lid for sealing the container for storing the graft, the lid having a convex portion that protrudes into the space inside the container when attached to the container, and a substantially annular groove on the underside of the convex portion, the lid being capable of pushing out gas and liquid from the container storing the graft and liquid to make it liquid-tight, and the groove on the underside being capable of capturing air bubbles in the liquid. [2] The transport container according to [1], wherein the lower surface of the convex portion is a flat surface parallel to the liquid surface in the container. [3] The transport container according to [1] or [2], wherein the side of the convex portion is configured to be in close contact with the inner surface of the container. [4] The transport container according to any one of [1] to [3], wherein a substantially annular groove is provided along the periphery of the lower surface. [5] The transport container according to any one of [1] to [4], wherein the substantially annular groove has an inverted trapezoidal cross section.

[0011] [6] The transport container according to any one of [1] to [5], further comprising one or more cut grooves intersecting the substantially annular groove portion. [7] The transport container according to any one of [1] to [6], wherein the graft is a sheet-shaped cell culture. [8] A method for transporting a graft, comprising the steps of providing a transport container described in any one of [1] to [7], attaching a lid to a container containing the graft and a liquid, and transporting the graft. [9] A method for culturing a graft, comprising the steps of providing a transport container according to any one of [1] to [7], seeding cells into the container containing a culture medium, attaching a lid to the container, and culturing the graft in the container. [Effects of the Invention]

[0012] According to the present invention, the air bubbles remaining in the container can be trapped and the graft can be protected with a simple mechanism and simple operation, which offers great advantages in terms of workability and manufacturing costs. Furthermore, since the device can be attached to a general-purpose petri dish with one touch, there is no need to use a special container, making it highly versatile.

[0013] In particular, since the capture of air bubbles can be achieved by the simple task of vibrating the container, the graft can be reliably protected without relying on the operator's technique. Furthermore, even if air bubbles are discovered in the container during transport, they can be easily captured without removing the lid, minimizing the occurrence of contamination. [Brief explanation of the drawings]

[0014] [Figure 1] Fig. 1A is a conceptual diagram showing a transport container including a lid according to a first embodiment, and Fig. 1B is a conceptual diagram showing a state in which the lid is attached to the container. [Figure 2] FIG. 2 shows an embodiment in which the lid of the present invention is attached to a general-purpose petri dish. [Figure 3] FIG. 3 is a detailed view of the shipping container of FIG. DETAILED DESCRIPTION OF THE INVENTION

[0015] In the present invention, the term "graft" refers to a structure for transplantation into a living body, and particularly refers to a transplant structure containing viable cells as a component. Preferably, the graft is a transplant structure that does not contain any structures (e.g., scaffolds) other than viable cells and substances derived from viable cells. Examples of the graft of the present invention include, but are not limited to, sheet-shaped cell cultures, spheroids, cell aggregates, etc., and are preferably sheet-shaped cell cultures or spheroids, and more preferably sheet-shaped cell cultures.

[0016] In the present invention, the term "sheet-shaped cell culture" refers to a sheet-like structure in which cells are interconnected. The cells may be interconnected directly (including via cellular elements such as adhesion molecules) and / or via an intervening substance. The intervening substance is not particularly limited as long as it can at least physically (mechanically) connect the cells, and examples thereof include extracellular matrix. The intervening substance is preferably derived from cells, particularly from the cells that constitute the cell culture. The cells are at least physically (mechanically) connected, but may also be functionally connected, for example, chemically or electrically. The sheet-shaped cell culture may be composed of one cell layer (single layer) or two or more cell layers (laminated (multilayered) structure, e.g., two, three, four, five, six, etc.). The sheet-shaped cell culture may also have a three-dimensional structure with a thickness exceeding that of a single cell, without the cells exhibiting a distinct layer structure. For example, in a vertical cross section of a sheet-shaped cell culture, cells may not be uniformly aligned in the horizontal direction, but may be arranged unevenly (eg, in a mosaic pattern).

[0017] The sheet-shaped cell culture preferably does not include a scaffold (support). Scaffolds are sometimes used in the art to attach cells to their surface and / or their interior and maintain the physical integrity of the sheet-shaped cell culture; for example, membranes made of polyvinylidene difluoride (PVDF) are known. However, the sheet-shaped cell culture of the present invention can maintain its physical integrity even without such a scaffold. Furthermore, the sheet-shaped cell culture of the present invention preferably consists only of substances derived from the cells that constitute the sheet-shaped cell culture, and does not include any other substances.

[0018] The cells constituting the sheet-shaped cell culture are not particularly limited as long as they can form a sheet-shaped cell culture, and include, for example, adhesive cells (adherent cells). Adherent cells include, for example, adherent somatic cells (e.g., cardiomyocytes, fibroblasts, epithelial cells, endothelial cells, hepatocytes, pancreatic cells, kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, synovial cells, chondrocytes, etc.) and stem cells (e.g., myoblasts, tissue stem cells such as cardiac stem cells, embryonic stem cells, pluripotent stem cells such as iPS (induced pluripotent stem) cells, mesenchymal stem cells, etc.). Somatic cells may also be differentiated from stem cells, particularly iPS cells (iPS cell-derived adherent cells). Non-limiting examples of cells that can be used to form sheet-shaped cell cultures include myoblasts (e.g., skeletal myoblasts), mesenchymal stem cells (e.g., derived from bone marrow, adipose tissue, peripheral blood, skin, hair roots, muscle tissue, endometrium, placenta, and umbilical cord blood), cardiomyocytes, fibroblasts, cardiac stem cells, embryonic stem cells, iPS cells, synovial cells, chondrocytes, epithelial cells (e.g., oral mucosal epithelial cells, retinal pigment epithelial cells, nasal mucosal epithelial cells), endothelial cells (e.g., vascular endothelial cells), hepatocytes (e.g., hepatic parenchymal cells), pancreatic cells (e.g., pancreatic islet cells), kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, and skin cells. Non-limiting examples of iPS cell-derived adhesive cells include iPS cell-derived cardiomyocytes, fibroblasts, epithelial cells, endothelial cells, hepatocytes, pancreatic cells, kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, synovial cells, and chondrocytes.

[0019] In the present invention, the container is not particularly limited as long as it can accommodate a graft, liquid, etc., and does not leak liquid, and any container, including commercially available containers, can be used. Examples of container materials include, but are not limited to, polyethylene, polypropylene, Teflon (registered trademark), polyethylene terephthalate, polymethyl methacrylate, nylon 6,6, polyvinyl alcohol, cellulose, silicone, polystyrene, glass, polyacrylamide, polydimethylacrylamide, and metals (e.g., iron, stainless steel, aluminum, copper, and brass). Furthermore, the container preferably has at least one flat bottom surface to maintain the shape of the graft, and examples thereof include, but are not limited to, petri dishes, cell culture dishes, and cell culture bottles. The area of ​​the flat bottom surface is not particularly limited, but is typically 1.13 to 78.5 cm. 2 , preferably 12.6 to 78.5 cm 2 , more preferably 9.1 to 60.8 cm 2 is.

[0020] In the present invention, the liquid in the container is composed of at least one component, and the component is not particularly limited, but may be, for example, water, an aqueous solution, a non-aqueous solution, a suspension, an emulsion, or the like. The liquid or fluid in the present invention may be any fluid that has overall fluidity, and may contain solid substances such as cell scaffolds and other non-liquid components such as air bubbles.

[0021] The components constituting the liquid in the container are not particularly limited as long as they have little effect on the graft. When the graft is a membrane made of a biomaterial, the components constituting the liquid in the container are preferably biocompatible from the viewpoint of biological stability and long-term storage potential, i.e., components that do not cause undesired effects such as inflammatory reactions, immune reactions, or toxic reactions in biological tissues or cells, or at least have minimal such effects. Examples of suitable components include water, physiological saline, physiological buffer solutions (e.g., HBSS, PBS, EBSS, Hepes, sodium bicarbonate, etc.), culture media (e.g., DMEM, MEM, F12, DMEM / F12, DME, RPMI164), and the like. 0, MCDB, L15, SkBM, RITC80-7, IMDM, etc.), sugar solutions (sucrose solution, Ficoll-paque (registered trademark) PLUS, etc.), seawater, serum-containing solutions, Renografin (registered trademark) solution, metrizamide solution, meglumine solution, glycerin, ethylene glycol, ammonia, benzene, toluene, acetone, ethyl alcohol, benzol, oil, mineral oil, animal fat, vegetable oil, olive oil, colloidal solution, liquid paraffin, turpentine oil, linseed oil, castor oil, etc.

[0022] When the graft is a sheet-shaped cell culture, the components constituting the liquid in the container are preferably capable of stably preserving the cells, contain the minimum amount of oxygen and nutrients necessary for cell survival, and do not destroy the cells due to osmotic pressure or the like. Examples of such liquids include, but are not limited to, physiological saline, physiological buffer solutions (e.g., HBSS, PBS, EBSS, Hepes, sodium bicarbonate, etc.), culture media (e.g., DMEM, MEM, F12, DMEM / F12, DME, RPMI1640, MCDB, L15, SkBM, RITC80-7, IMDM, etc.), sugar solutions (sucrose solution, Ficoll-paque PLUS (registered trademark), etc.), etc.

[0023] The amount of liquid in the container is not particularly limited as long as it is sufficient to hold the graft with the lid attached to the container, and the liquid volume formed between the bottom surface of the container and the underside of the lid is high enough to prevent the graft from shaking. In one embodiment of the present invention, the sheet-shaped cell culture has a diameter of approximately 35 to 55 mm and an area of ​​6 cm. 2The liquid volume is, for example, 1.0 mm to 70.0 mm, regardless of the diameter of the sheet-shaped cell culture.

[0024] In the present invention, the lid is not particularly limited as long as it seals the container. Examples of materials for the lid include, but are not limited to, polyethylene, polypropylene, Teflon (registered trademark), polyethylene terephthalate, polymethyl methacrylate, nylon 6,6, polyvinyl alcohol, cellulose, silicone, polystyrene, glass, polyacrylamide, polydimethylacrylamide, and metals (e.g., iron, stainless steel, aluminum, copper, and brass).

[0025] In the present invention, the shapes of the lid and the container are not particularly limited as long as the lid and the container can be engaged with each other and such engagement can form a sealed space. For example, when the container is a general-purpose petri dish, it is preferable that the lid be circular. Furthermore, the lid and / or the container may be made of a light-transmitting material so that the condition of the graft contained in the container and the presence or absence of air bubbles in the liquid can be confirmed.

[0026] In the present invention, the "space within the container" refers to the space below the opening of the container, i.e., the storage space of the container that can store liquid, gas, grafts, etc. Additionally, the "attached to the container" refers to the state in which the lid is attached to the container and the space within the container is sealed.

[0027] In the present invention, the "protrusion" refers to a portion that protrudes downward from the top plate of the lid. That is, the protrusion refers to a portion that protrudes into the space within the container when the lid is attached to the container and can push out gas and liquid from the container. The protrusion has a shape that can fit into the space above the liquid level within the container to achieve liquid-tightness. For example, if the container to be used is a circular, general-purpose Petri dish, the protrusion has a cylindrical shape, and the side of the protrusion is sized to fit closely to the side of the general-purpose Petri dish. The protrusion may be hollow or solid. If the protrusion is hollow, the cross section of the lid will be concave, and by making the underside of the protrusion optically transparent, it becomes easier to check the condition of the graft contained in the container and the location of air bubbles in the liquid.

[0028] In the present invention, the "lower surface of the convex portion" refers to the lowermost surface that first comes into contact with the liquid surface when the lid is attached to the container. That is, the convex portion has a three-dimensional shape with side surfaces and a lower surface, and is dimensioned so that the space inside the container when the lid is attached to the container is filled with the convex portion and the liquid. Furthermore, the lower surface of the convex portion is a flat surface parallel to the liquid surface in the container, and is configured to suppress the generation of water flow when the convex portion comes into contact with the liquid surface. The lower surface of the convex portion is flat except for the groove portion. Conversely, for example, if the horizontal cross-sectional area of ​​the convex portion increases (or decreases) as it goes vertically downward, water flow will occur when the convex portion enters the liquid, which may cause wrinkles, tears, damage, etc. to the graft in the liquid.

[0029] In the present invention, the term "groove" refers to a concave groove carved into the underside of the convex portion toward the top plate of the lid. In other words, bubbles remaining in the space within the container tend to rise toward the top plate of the lid, and are therefore suitably captured by the groove carved toward the top plate. Furthermore, the term "substantially annular groove" includes both continuous and discontinuous grooves extending in the circumferential direction of the underside. In other words, bubbles present on the underside are surrounded by the substantially annular groove, and can be easily captured by the groove simply by vibrating (e.g., tilting) the container. Furthermore, for example, bubbles that rise to the surface can also be naturally captured by the groove simply by leaving the container as is without vibration.

[0030] To ensure proper trapping of air bubbles, the groove is preferably provided on the periphery of the lower surface. This allows air bubbles that may remain on the lower surface to be surrounded, making trapping easier. The cross section of the groove is not limited to a square, circle, trapezoid, inverted trapezoid, etc., but an inverted trapezoid is preferred to prevent trapped air bubbles from returning to the space within the container. The width of the groove can be 0.1 mm to 3.0 mm, more preferably 0.8 mm to 1.3 mm.

[0031] A transport container according to a preferred embodiment of the present invention will be described in detail below with reference to the drawings. In this embodiment, the container is a circular general-purpose petri dish, and the lid has a cylindrical protrusion that fits into the space inside the container of the general-purpose petri dish. Fig. 1A is a conceptual diagram showing a transport container including a lid according to a first embodiment. Fig. 1B is a conceptual diagram showing the state in which the lid is attached to the container. Fig. 2 shows an example in which the lid of the present invention is attached to a general-purpose petri dish.

[0032] As shown in FIG. 1, a transport container according to a first embodiment of the present invention includes a lid 1 that seals a container C for containing a graft S. The lid 1 includes a top plate 11 and a convex portion 12 that protrudes downward from the top plate 11. As shown in FIG. 1A, the convex portion 12 is shaped like a solid cylinder having a side surface 13 and a bottom surface 14, and the side surface 13 has a slight draft angle to match the shape of the inner surface of a typical petri dish. The bottom surface 14 of the convex portion 12 is shaped as a flat surface parallel to the liquid surface so as to suppress the generation of a water flow when the convex portion 12 comes into contact with the liquid surface.

[0033] As shown in the bottom of Fig. 1A, the bottom surface 14 has a circular shape, and is provided with an annular groove 15 along its periphery. Six cut grooves 17 are provided in the groove 15 in a direction intersecting the groove 15. The six cut grooves 17 are arranged at approximately equal intervals in the circumferential direction of the groove 15. Two opposing notches 18 are provided on the edge of the top plate 11. An annular recess 16 is provided on the top plate 11 to surround the protrusion 12 in order to receive the opening edge of the container C (top of Fig. 1A).

[0034] FIG. 1B shows a conceptual diagram of a lid 1', a modified version of the lid 1 in FIG. 1A, attached to a container C. As described above, the convex portion 12 of the lid 1 is formed in a solid cylindrical shape, while the convex portion 12 of the lid 1' is formed in a hollow cylindrical shape. That is, the cross section of the lid 1' is concave, and the underside 14 is thinner than that of the lid 1, making it easier to visually check air bubbles and the state of the graft S inside the container C. The top plate 11 is attached to the edge of the container C and is configured to seal the edge of the container C. The top plate 11 decreases in height in two stages toward the center of the container. The first stage, which exists from the top plate 11 toward the center, is an annular flat surface, and the second stage is a circular flat surface. The first stage exists directly above the groove portion 15, and the second stage forms a gas-permeable membrane 19. The gas permeable membrane 19 is configured so that, with the lid 1' attached to the container C, air necessary for culturing the graft S and maintaining its quality can be supplied into the container C.

[0035] As shown in FIG. 1B, when attaching the lid 1' to the container C, first, the liquid L and the graft S are placed in the container C. At this time, the liquid level of the liquid L in the container C is adjusted so that when the container C is sealed with the lid 1', the liquid level is approximately the same height as the lower surface 14 of the lid 1' or slightly higher than the lower surface 14. Next, the lid 1' is attached to the container C, and the gas and liquid L in the space inside the container C are pushed out by the lid 1' (protrusion 12) to make it liquid-tight. By adjusting the liquid level as described above, excess liquid L is prevented from being pushed out.

[0036] The protrusion 12 of the lid 1' is molded to fit into the space above the liquid level within the container C, so the side surface 13 of the protrusion 12 adheres closely to the inner surface of the container C. This ensures that any remaining air bubbles B are present on the flat lower surface 14, making them easy to visually check. Furthermore, because the lower surface 14 of the lid 1' is a flat surface parallel to the liquid surface, it is possible to minimize water currents that occur when the graft S comes into contact with or leaves the liquid surface. This is particularly advantageous when the graft S is a sheet-shaped cell culture, which is extremely fragile.

[0037] When the lid 1' is attached to the container C, air pockets are formed in the annular groove 15 and the cut groove 17 (although they may not be formed). If air bubbles B remaining on the underside 14 of the lid 1' are found after the attachment work is completed, slightly tilt the container C (lid 1') to move the air bubbles B in the radial direction and cause them to adhere to the air pockets. When tilting the container C, insert two fingers (for example, the thumb and middle finger) into the notches 18 from above the top plate 11 and lift the container C while pinching it, which prevents unnecessary force from being applied to the top plate 11 and causing it to come off.

[0038] The air bubbles B adhering to the air reservoirs are integrated (trapped) with the air in the grooves 15, and are prevented from moving toward the center where the graft S is present. Even when no air reservoirs are formed, i.e., when the grooves 15 are filled with the liquid L, the air bubbles B easily enter and are trapped in the grooves 15. The grooves 15 are provided along the periphery of the lower surface 14, i.e., so as to surround the air bubbles B on the lower surface 14, so that the air bubbles B can be integrated with the air in the grooves 15 regardless of the direction in which the container C (lid 1') is tilted.

[0039] The cut grooves 17 serve to block the movement of the bubbles B in the circumferential direction when the container C is tilted left or right and the bubbles B move along the groove portions 15. Furthermore, while the groove portions 15 are linear grooves, the cross grooves where the groove portions 15 and the cut grooves 17 intersect are planar grooves, and therefore can more effectively capture the bubbles B. By making the cross section of the groove portions 15 an inverted trapezoid, it is also possible to make it difficult for air (air pockets, bubbles B) to move downward. The opening edge of the container C fits into the annular recess 16 of the lid 1', making it difficult for it to come off.

[0040] Another aspect of the present invention relates to a method for transporting an implant, the method comprising the steps of providing a transport container of the present invention, attaching a lid to a container containing an implant and a liquid, and transporting. The graft is transferred by placing the graft and liquid in a container with a lid attached. Air bubbles are captured by moving them into the groove beforehand through vibration during transfer, or by tilting the container to move the bubbles into the groove, allowing the graft to be transferred in a protected state. The transfer method may include a step of capturing air bubbles in the groove (vibrating the container).

[0041] Another aspect of the present invention relates to a method for culturing an explant, the method comprising the steps of providing a transport container of the present invention, seeding cells into the container containing a culture medium, attaching a lid to the container, and culturing an explant in the container.

[0042] The step of seeding cells into a container containing medium can be achieved, for example, by filling the UpCell with 10% FBS-DMEM / F12 medium and seeding cells at 2.2 x 10e7 cells / 9 mL when the graft is a sheet-shaped cell culture. The step of attaching a lid to the container can also be performed after cell culture has progressed to a certain extent. The step of culturing the graft in the container includes culturing the graft with the lid attached to the container, and can be achieved, for example, by placing the container in an incubator or supplying air through a gas-permeable membrane.

[0043] For example, the initial steps of the culture method, i.e., providing a transport container, seeding cells into the container containing the culture medium, and attaching the lid to the container, can be performed in a clean room, and subsequent steps can be performed outside the clean room. This allows for more flexibility in the transport and manufacturing processes, as air bubbles found outside the clean room can be captured without removing the lid, and explant culture can be performed outside the clean room.

[0044] Furthermore, currently, sheet-shaped cultures are peeled off from containers after culturing is completed and transferred to transfer containers, which often results in damage to the sheet-shaped cultures. However, the present invention allows the processes from culturing to transfer to be carried out in the same container, minimizing damage to the sheet-shaped cultures and eliminating the need for transfer. Furthermore, even if air bubbles are discovered during culturing or transfer, they can be captured without removing the lid. The culture method may include a step of capturing the air bubbles in the grooves.

[0045] As described above, the present invention has a simple mechanism and can capture residual air bubbles in a container and protect a graft with a simple operation, which offers great advantages in terms of workability and manufacturing costs. Furthermore, it can be attached to a general-purpose petri dish with one touch, eliminating the need for a special container, making it highly versatile.

[0046] In particular, since the capture of air bubbles can be achieved by the simple task of vibrating the container, the graft can be reliably protected without relying on the operator's technique. Furthermore, even if air bubbles are discovered in the container during transport, they can be easily captured without removing the lid, minimizing the occurrence of contamination.

[0047] Although the present invention has been described with reference to the illustrated embodiment, the present invention is not limited to this embodiment. In the present invention, each component can be replaced with any component that can perform the same function, or any component can be added.

[0048] Example A physiological buffer solution was placed in a general-purpose Petri dish (Thermo Fisher, 6 cm upcell) to a liquid volume of 5 mm, and a sheet-shaped cell culture was placed in the liquid. The transport container (lid) of the present invention was then attached to the general-purpose Petri dish. Remaining air bubbles were confirmed, so the container was tilted to guide the bubbles into the groove. As shown in Figure 2A (photographed from the bottom of the container) and Figure 2B (photographed from the side of the container), the air bubbles (arrowheads) were trapped in the groove, and did not move to the center even when the container was tilted left or right. [Explanation of symbols]

[0049] C container L liquid S graft B. Air bubbles 1, 1' Lid (transport container) 11 Top plate 12 Convex part 13 Side 14 Bottom side 15 Groove 16 Recess 17 Cutting groove 18 Cutout 19 Gas-permeable membrane

Claims

1. A transport container for protecting a graft, comprising a lid that seals the container for storing the graft, the lid having a convex portion that protrudes toward the space inside the container when attached to the container, and a substantially annular groove on the underside of the convex portion, the lid being able to push out gas and liquid from the container storing the graft and liquid, making it liquid-tight, and the groove on the underside being able to capture air bubbles in the liquid.

2. 2. The transport container according to claim 1, wherein the lower surface of the convex portion is a flat surface parallel to the liquid surface in the container.

3. 3. The transport container according to claim 1, wherein the side surface of the protrusion is configured to be in close contact with the inner surface of the container.

4. The transport container according to any one of claims 1 to 3, wherein a substantially annular groove is provided along the periphery of the lower surface.

5. The transport container according to any one of claims 1 to 4, wherein the substantially annular groove has an inverted trapezoidal cross section.

6. The shipping container according to any one of claims 1 to 5, further comprising one or more cut grooves intersecting the generally annular groove portion.

7. The transport container according to any one of claims 1 to 6, wherein the graft is a sheet-shaped cell culture.

8. A method for transporting an implant, comprising the steps of providing a transport container according to any one of claims 1 to 7, attaching a lid to a container containing the implant and a liquid, and transporting.

9. A method for culturing an explant, comprising the steps of providing a transport container according to any one of claims 1 to 7, seeding cells into the container containing a culture medium, attaching a lid to the container, and culturing the explant in the container.

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