JP2018121568A - Container for transporting sheet-like biological tissue and method of transporting same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a container and a method thereof for preventing damage to a sheet-like biological tissue due to vibration and metabolism inhibition. The present invention is a container for storing and / or transporting a sheet-shaped living tissue, and a storage space for accommodating the sheet-shaped living tissue and a preservative solution is formed by a lower bottom portion of the main body and a lower side portion of the main body. The lower part of the main body and the upper part of the lower side of the main body are placed to seal the accommodation space, and the upper part of the main body having at least one air hole and the air hole are sealed to seal the gas permeability. A member is included, wherein at least a part of the lower bottom of the main body is provided with a lower convex portion, and here, at least a part of the inner surface of the upper part of the upper part of the main body is provided with an upper convex portion. When combined with the upper part of the main body, the gap between the lower convex portion and the upper convex portion provides a container capable of holding the sheet-like biological tissue in a predetermined space in the accommodation space. [Selection diagram] FIG. 4A

Description

  The present invention relates to a container for transporting a sheet-like living tissue. The present invention also relates to a method for transporting a sheet-like living tissue.

  In recent years, in the field of regenerative medicine, a method for regenerating an organ or tissue having damage or disease has been sought. Attempts have been made to promote regeneration of organs and tissues by directly applying therapeutic substances to these organs and tissues. As one of the regenerative medicine technologies, a technology has been developed that uses a culture dish coated with a temperature-responsive polymer and collects cells as a therapeutic substance in the form of a sheet. (Patent Documents 1 to 3). This technique has made it possible to transplant sheet-like cultured cells.

  Conventionally, when transplanting such sheet-shaped cultured cells to an affected area, it is necessary to transport the culture dishes to which the cultured cells are attached one by one to the transplantation site and collect them as sheet-shaped cultured cells immediately before transplantation. In addition, in order to increase the number of cells to be transplanted, when it is necessary to stack the sheet-like cultured cells, it is necessary to perform a stacking process in the field. However, the layering process at the transplantation site has many problems such as (1) time-consuming, (2) skillful technology required, (3) difficult to process while maintaining sterility. It was.

  Therefore, a method has been devised in which a sheet-like cultured cell is produced by a skilled technician and transported to a transplantation site in a facility where sterility is maintained, not in the transplantation site. Transport containers used in this method have been developed and disclosed in, for example, Patent Documents 4 to 6.

Japanese Patent Laid-Open No. 02-21865 JP 05-192138 A International Publication No. 2002/008387 JP 2002-335950 A JP 2007-119033 A JP 2004-187518 A

Conventional containers for transporting sheet-like living tissues are not able to interfere with the metabolism of cultured tissues to impair biological activity, break the sheet-like living tissues due to physical vibration, or vibration during transportation. There have been problems such as leakage of the preservation solution and foaming of the preservation solution by exposure to low atmospheric pressure during air transportation. For example, the containers disclosed in Patent Documents 4 and 5 have a system in which a cultured tissue is sandwiched and held by a support member, so that the metabolism of the cells is hindered and the cultured tissue cannot withstand long-term storage and transportation. The container disclosed in Patent Document 6 is not necessarily configured to be wrapped with a support member. However, since the cultured tissue is sealed in a narrow space between the tray and the lid, metabolism is hindered, and storage and transportation for a long time is performed. However, there is a problem that the cultured tissue cannot tolerate. In particular, the cultured tissue cannot remove gas such as carbon dioxide generated by metabolism, and gas such as oxygen necessary for metabolism cannot be supplied, so that metabolism is hindered.
The present invention aims to solve such problems.

  The present invention solves the above problems. That is, the present invention has, for example, the following configurations / features.

[1] A container for storing and / or transporting a sheet-like living tissue,
A lower part of the main body in which a housing space for accommodating the sheet-like living tissue and the preservation solution is formed by the lower part of the main body and the side of the lower part of the main body
An upper part of the main body which is placed on the upper part of the lower part of the main body to seal the accommodation space and has at least one air hole;
A gas-permeable sealing member for sealing the air hole;
Including
Here, at least a part of the bottom portion of the main body includes a lower convex portion,
Here, an upper convex portion is provided on at least a part of the main body upper inner surface of the main body,
When the lower part of the main body and the upper part of the main body are combined, the gap between the lower convex part and the upper convex part can hold the sheet-like living tissue in a predetermined space in the accommodation space. .
[2] The container according to [1], wherein the lower convex portion includes at least one lower groove.
[3] The container according to [1] or [2], wherein the upper convex portion includes at least one upper groove.
[4] The container according to any one of [1] to [3], wherein the inner surface of the upper part of the main body has a slope toward the air hole.
[5] The container according to any one of [1] to [4], wherein the sealing member is a liquid-impermeable sealing film.
[6] The container according to any one of [1] to [5], wherein the sheet-like living tissue further includes a support.
[7] The container according to [6], wherein the support has a shape that supports only a peripheral region of the sheet-like living tissue.
[8] The container according to any one of [1] to [6], wherein the upper part of the main body and / or the lower part of the main body includes a collar part.

[9] A method for storing and / or transporting a sheet-like living tissue,
(1) A sheet-like living tissue is placed on the lower convex portion of the lower part of the main body constituting the container according to any one of [1] to [8],
(2) Apply the upper part of the main body to the lower part of the main body,
(3) Fill the storage solution from the air hole to eliminate the gas,
(4) plug the air hole with a sealing member;
(5) storing and / or transporting the container on which the sheet-like living tissue is placed;
Including a method.
[10]
The method according to [9], wherein the sheet-like living tissue includes a support.

  According to the container and the method using the container of the present invention, it is possible to prevent damage to the sheet-like living tissue and liquid leakage from the container due to vibration during storage and transportation of the sheet-like living tissue. In addition, the sheet-like living tissue can be stored and / or transported while maintaining the quality of the sheet-like living tissue, and as a result, it is possible to provide a sheet-like living tissue that can be used quickly, simply, and reliably at the site of surgery or the like. It becomes.

It is a top view which shows the container of this invention. It is a front view which shows the container of this invention. It is a top view which shows the main body lower part which comprises the container of this invention. It is a perspective view which shows the main body lower part which comprises the container of this invention. It is a front view which shows the main body lower part which comprises the container of this invention. It is a front view which shows the main body lower part which comprises the container of this invention. It is a bottom view which shows the main body upper part which comprises the container of this invention. It is a perspective view which shows the main body upper part which comprises the container of this invention. It is a front view which shows the main body upper part which comprises the container of this invention. It is a schematic diagram which shows the usage example of the container of this invention. It is a schematic diagram which shows the usage example of the container of this invention. It is a schematic diagram (cross section) which shows the usage example of the container of this invention. It is a schematic diagram (cross section) which shows the usage example of the container of this invention. It is a top view which shows the main body lower part which comprises the container of this invention (modification). It is a figure which shows the container of this invention (modified example). It is a figure which shows the container of this invention (modified example).

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. The configuration of the embodiment is an exemplification, and the configuration of the present invention is not limited to the specific configuration of the following embodiment.

(Configuration example of container for storage and / or transportation of sheet-like biological tissue)
1A (plan view) and FIG. 1B (front view) are diagrams showing a configuration example of a storage and / or transportation container (also simply referred to as “container 1”) for sheet-like biological tissue according to an embodiment of the present invention. It is. The container 1 covers a main body lower part 2 for accommodating a sheet-like living tissue 5 (see FIGS. 4A to 4C) described later, and the main body lower part 2, and an accommodation space S (see FIGS. 2A and B) of the main body lower part 2. A main body upper part 3 as a lid for sealing, and a sealing member 6 for sealing an air hole 35 provided in the main body upper part 3 are included. Examples of the material of the lower body 2 and the upper body 3 include, for example, polyethylene, polypropylene, polycarbonate, polystyrene, polyvinyl chloride, nylon, polyurethane, polyurea, polylactic acid, polyglycolic acid, polyvinyl alcohol, polyvinyl acetate, and poly (meth). Acrylic acid, poly (meth) acrylic acid derivative, polyacrylonitrile, poly (meth) acrylamide, poly (meth) acrylamide derivative, polysulfone, polycarbonate, cellulose, cellulose derivative, polysilicon, glass, ceramic, metal and the like can be mentioned.

  The “sheet-like biological tissue” stored and / or transported by the container of the present invention and the method using the same is a very thin tissue, and contracts when not attached to a support and is difficult to handle. Further, in a state where the sheet-like biological tissue is suspended in the preservation solution, there is a problem in that the sheet-like biological tissue is aggregated due to the adhesive component of the sheet-like biological tissue. Therefore, when moving the sheet-like biological tissue, it is handled in a state of being attached to the “support”. Depending on the type of “sheet-like living tissue”, there are some that are extremely thick and have a certain degree of rigidity, and there are some that can be handled without the need for a “support”. Now, an example in which the “sheet-like biological tissue” attached to the “support” is accommodated will be described.

(Example configuration of the bottom of the main unit)
2A to D show a configuration example of the lower main body 2 constituting the container 1 according to the embodiment of the present invention. In the present embodiment, the main body lower portion 2 includes a main body lower portion 21 and a hollow cylindrical main portion lower portion 22 provided on the lower portion 21 of the main body. A housing space S for housing the sheet-like living tissue 5 and the preservation solution 7 (see FIG. 4D) is formed by the lower body bottom portion 21 and the lower body side portion 22. A part of the peripheral region of the sheet-like living tissue 5 is placed on at least a part of the lower body bottom portion 21 (see FIGS. 4A to 4C), and the sheet-like living tissue 5 is separated from the bottom lower portion 21 of the body. The lower convex part 23 is provided.

  In the present embodiment, the lower convex portion 23 is disposed along the circumference of the main body lower bottom portion 21 and in contact with the side inner wall surface 22a. A lower groove 23b is provided between adjacent lower protrusions 23. By providing the lower groove 23b, a jig such as tweezers can be inserted. Thereby, the sheet-like biological tissue 5 can be easily taken out from the main body lower part 2 at the time of use.

As in the present embodiment, a plurality of lower protrusions 23 may be provided by providing lower grooves 23b at regular intervals, or may be a hollow cylindrical shape (donut shape) without providing lower grooves 23b (not shown). ). The lower groove 23b allows the metabolic gas diffused in the storage solution in the lower chamber 28 to escape to the upper chamber 38 by allowing the lower chamber 28 and the upper chamber 38 to communicate with each other, and oxygen in the upper chamber 38 is allowed to escape. It is preferable to provide one or more. In addition, the provision of the lower groove 23b also has an effect that the sheet-like living tissue 5 can be easily taken out by a jig such as tweezers during use.

  In one embodiment, when a plurality of lower protrusions 23 are provided, the lower protrusions 23 may be disposed directly below an upper protrusion 33 described later, or may be disposed directly below the upper groove 33b ( For example, see the cross-sectional views of FIGS. 4C and D). In another embodiment, the lower convex portion 23 of the main body lower portion 2 may be a hollow cylindrical shape not provided with the lower groove 23b (not shown).

  In the lower convex portion 23, the surface on which the sheet-like living tissue 5 is placed (referred to as “mounting surface 23a”) is preferably on the same plane. Thereby, the sheet-like biological tissue 5 placed on the lower convex portion 23 can be kept flat. In addition, the vertical height of the lower convex portion 23 is determined in relation to the thickness of the sheet-like living tissue 5 and / or the shape, size, or arrangement of the upper convex portion 33 (see FIGS. 3A and B). The The shape of the lower convex part 23 and the shape of the mounting surface 23a are not limited as long as the sheet-like living tissue 5 is separated from the main body lower bottom part 21 and is kept flat. Examples of the shape of the lower protrusion 23 include a columnar shape, a truncated cone shape, and a prismatic shape. Moreover, the shape of the lower convex part 23 may be a hollow cylindrical shape or a hollow prismatic shape. The larger the placement surface 23a, the higher the effect of stably keeping the sheet-like living tissue 5 flat.

  The arrangement of the lower protrusions 23 in the main body lower bottom 21 is also determined by the relationship between the shape of the container 1 and the shape of the desired sheet-like living tissue 5. For example, when the substantially circular sheet-like biological tissue 5 as in the present embodiment is accommodated, it is arranged at a position that supports the peripheral region of the sheet-like biological tissue 5, for example, the circumference of the sheet-like biological tissue 5. For example, when the sheet-like living tissue 5 is a polygon (a triangle, a quadrangle, a pentagon, a hexagon, etc.), it is arranged at a position that supports each corner and each side (see FIGS. 5A and B). Thereby, the sheet-like biological tissue 5 can be separated from the main body lower bottom part 21, and it can reduce that the sheet-like biological tissue 5 is damaged by the vibration which arises at the time of transport.

  In one embodiment of the present invention, the side outer wall surface 22 b of the main body lower side portion 22 may include an O-ring 26. The O-ring 26 is attached to an O-ring groove 22c provided on the side outer wall surface 22b. Due to the presence of the O-ring 26, when the main body upper part 3 is fitted to the main body lower part 2, fluid and / or gas flows in and out between the side outer wall surface 22b and the main body upper side inner wall surface 32a (see FIG. 3B). Can be prevented. The O-ring 26 is not limited as long as it is a member that seals the space between the main body lower portion 2 and the main body upper portion 3. As a material for the O-ring 26, for example, nitrile rubber, silicone rubber, fluoro rubber, urethane rubber, butyl rubber, polytetrafluoroethylene, metal, or the like can be used. The O-ring is not limited to the above as long as the same effect can be obtained.

  In another embodiment of the present invention, the container 1 may be provided with a male screw structure on the side outer wall surface 22b and a female screw structure on the main body upper side inner wall surface 32a instead of the O-ring 26 ( Not shown). By screwing the main body upper portion 3 into the main body lower portion 2, it is possible to prevent fluid and / or gas from flowing in and out between the side outer wall surface 22b and the main body upper side wall surface 32a.

  In the embodiment of the present invention, the main body lower part 2 is provided with a main body lower collar part 27 on the outer edge of the main body lower part bottom part 21. The main body lower collar part 27 is larger than the area of the main body lower bottom part 21, and helps to keep the main body lower part 2 horizontal. The main body lower collar part 27 is not essential, but when the container 1 is relatively small, for example, smaller than the diameter of a commercially available 35 mm culture dish, the container 1 falls down by having the main body lower collar part 27. Can be prevented. The size and shape of the main body lower collar 27 may be appropriately modified according to the purpose, and is not limited to this embodiment.

(Configuration example of the upper part of the main unit)
3A to 3C show a configuration example of the main body upper part 3 constituting the container 1 in one embodiment of the present invention. FIG. 3A is a bottom view of the main body upper part 3 as viewed from the main body lower part 2 side. FIG. 3B is a perspective view of the main body upper portion 3. 3C is a side view of the upper part 3 of the main body as viewed from the side, and the lower side of the drawing is the side applied to the lower part 2 of the main body. The main body upper part 3 has an action of limiting a movable range so that the support 4 to which the sheet-like living tissue 5 is attached does not move greatly due to vibration or the like. In addition, an operation of venting gas can be performed using the air slope 37 and the air hole 35 provided in the upper part 3 of the main body.

  In the present embodiment, the main body upper portion 3 includes a main body upper ceiling portion 31 having an air hole 35 and a main body upper side portion 32 provided on the main body upper ceiling portion 31. The main body upper part 3 is placed on the upper part of the main body lower side part 22 of the main body lower part 2 and seals the accommodation space S by combining with the sealing member 6 (see FIGS. 1A and 1B and FIG. 4D). The accommodation space S is divided into an upper chamber 38 and a lower chamber 28 by the sheet-like living tissue 5 supported by the support body 4 (see FIG. 4D). From the air hole 35, the storage space 7 can be filled into the accommodation space S. Moreover, the gas remaining in the accommodation space S can be eliminated by filling the storage liquid 7 from the air hole 35. The air hole 35 is sealed by a removable sealing member 6. As the sealing member 6, a sealing member 6 such as a plug type, a cap type, or a film type can be used, but preferably has a gas-permeable function. Examples of the plug-type sealing member 6 include natural rubber plugs, nitrile rubber plugs, silicone rubber plugs, fluorine rubber plugs, butyl rubber plugs, urethane plugs, silicone resin plugs (for example, silicosene (registered trademark), biosilico (registered) Trademark))) and the like. The plug-type sealing member 6 is preferably a urethane plug or a silicone resin plug having gas permeability. By using the sealing member 6 having gas permeability, it is possible to maintain gas exchange necessary for breathing even in the sheet-like living tissue 5 containing materials (for example, cells and living tissues) that require breathing. it can. Examples of the cap-type sealing member 6 include a silicone resin cap and a polypropylene cap. As the film-type sealing member 6, it is possible to use a gas permeable and liquid impermeable sealing film such as a polypropylene film, a rayon film, a polyurethane film, or the like. Examples of commercially available film-type sealing members 6 include Plate Seal AeroSeal (trademark) (EXCEL Scientific, CA, USA), Breeze Easy (Toho, Tokyo, Japan), Breathable Film (Corning, NY, USA). The material is not limited to the above example, but is not limited as long as the fluid can be prevented from leaking from the air hole 35 and is gas permeable and does not pass bacteria, viruses, or the like. Even when the accommodation space S is sealed by the sealing member 6, the living sheet-like living tissue is alive through the air holes 35 provided in the upper portion 3 of the main body and the gas-permeable sealing member 6. Oxygen necessary for this is continuously supplied from the outside of the container 1 to the preservation solution. Further, metabolic gas generated by metabolism of the sheet-like living tissue stored in the container 1 is also discharged out of the container 1 through the air hole 35 and the gas permeable sealing member 6.

  Moreover, when opening, the air hole 35 is provided in the main body upper part 3 of the container 1, and the sealing member 6 is provided, so that the main body upper part 3 is removed from the main body lower part 2 before the sealing member is removed. 6 can be removed. As a result, air flows into the accommodation space S from the air hole 35, and the pressure difference between the inside and outside of the container 1 is eliminated. As a result, the main body 3 can be easily and safely removed without damaging the sheet-like living tissue due to the intense flow of the preservation solution.

  The number of air holes 35 provided in the upper part 3 of the main body may be one, or two or more. When a plurality of air holes 35 are provided, the sealing member 6 needs to have a number that can seal all of the air holes 35. When the sealing member 6 is, for example, a film type, all the air holes 35 may be sealed with a single film type sealing member 6.

  The main body upper inner surface 31a includes an upper convex portion 33 that prevents the sheet-like living tissue 5 from coming into contact with the main body upper inner surface 31a. In one embodiment, the shape of the upper convex portion 33 is a hollow cylindrical shape having a portion (shown as “upper groove 33b” in FIGS. 3A and 3B) in which a part of the side surface is missing. The upper groove 33b allows the metabolic gas dissolved and diffused in the storage solution in the lower chamber 28 to escape by diffusing into the upper chamber 38 by allowing the lower chamber 28 and the upper chamber 38 to communicate with each other. The oxygen dissolved in the storage solution is diffused into the lower chamber 28. In another embodiment, it may be a hollow cylindrical upper convex portion 33 not provided with an upper groove 33b (not shown).

  The arrangement of the upper convex portion 33 is determined depending on the shape of the container 1 and the desired shape of the sheet-like living tissue 5. For example, when accommodating the substantially circular sheet-like living tissue 5 as in the present embodiment, the upper convex portion 33 is formed on the peripheral region of the sheet-like living tissue 5, for example, on the inner side of the circumference of the sheet-like living tissue 5. It is provided to be arranged.

  In order to prevent the sheet-like living tissue 5 from sticking to the main body upper inner surface 31a and damage due to vibration during transportation when the main body upper portion 3 is detached, the surface area of the tip surface of the upper convex portion 33 (upper convex portion lower surface 33a). Is preferably smaller. Particularly preferably, the upper convex portion 33 includes an upper convex portion lower surface 33a having an area smaller than the placement surface 23a. The upper convex portion lower surface 33a is preferably in the same plane. The shape of the upper convex portion 33 and the shape of the upper convex portion lower surface 33a are not limited as long as the sheet-like living tissue 5 can be attached to the main body upper inner surface 31a and can be prevented from being damaged by vibration during transportation. For example, examples of the shape of the upper convex portion 33 include a columnar shape, a truncated cone shape, and a prismatic shape.

  In one embodiment, the main body upper inner surface 31a preferably has a slope toward the air hole 35 (“air slope 37” in FIGS. 4C and D). Thereby, when the container 1 is filled with the preservation solution 7, the gas (bubbles) contained in the container 1 (the accommodation space S) is easily guided toward the air hole 35. By reducing the gas contained in the container 1, the opportunity for bubbles to damage the sheet-like living tissue 5 is reduced. The air slope 37 may be conical or pyramidal. In another embodiment, the air hole 35 may further have an opening such as a hollow cylindrical shape or a hollow prism shape communicating with the air hole 35 (not shown).

  In one embodiment, the main body upper portion 3 may have a main body upper flange portion 36. The main body upper collar part 36 facilitates the attachment and detachment of the main body upper part 3 to the main body lower part 2. The main body upper collar part 36 is not limited as long as the main body upper part 3 can be easily attached and detached.

  In one embodiment, the main body upper part 3 has a key 34 for fixing the main body upper part 3 to an appropriate position of the main body lower part 2. In the present embodiment, the key 34 is formed integrally with the upper convex portion 33. The key 34 fits in the key groove 24 of the lower body 2 and fixes the upper body 3 in an appropriate position. Moreover, the main body 3 is prevented from rotating with respect to the main body lower part 2 by fitting the key 34 into the key groove 24. The key 34 and the key groove 24 may have any shape as long as the main body upper portion 3 does not rotate, and the shape and arrangement thereof are not limited.

(Configuration example when using a container for storing and / or transporting a sheet-like living tissue)
4A to 4D show configuration examples when the container 1 of the present invention is used. The sheet-like living tissue 5 is placed on the lower convex portion 23 of the lower body 2 and the upper body 3 is applied to the lower body 2 (see FIGS. 4A and B). Thereafter, the preservation solution 7 is filled from the air hole 35 in the upper part 3 of the main body. At this time, by having the air slope, the bubbles 9 move to the air holes 35 and are excluded from the container 1 (see FIG. 4C). Thereafter, the air hole 35 is closed with the sealing member 6 (see FIG. 4D). Thereby, it becomes possible to preserve | save and / or transport the container 1 in which the sheet-like biological tissue 5 is mounted.

  In one embodiment, a sheet-like living tissue 5 in close contact with the support 4 is used. For example, when the sheet-like living tissue 5 to be accommodated is a sheet-like cell group obtained by culturing cells (referred to as a “cell sheet”), the cell sheet peeled off from the culture substrate contracts, and the container Not suitable for use in 1. Therefore, in this embodiment, the cell sheet is preferably used together with the support 4. The support 4 used in this embodiment is preferably one that allows the storage solution to pass through gas generated by metabolism of cells and gas required for metabolism of cells (for example, has a mesh structure). The inner region of the circular support may have a concentric shape. An effect of not impeding metabolism is that the cell sheet directly touches the preservation solution by the defective part. In this case, when the cell sheet is brought into close contact, the edge portion of the cell sheet is supported, and the cell sheet inner side portion is not supported. Thereby, the cell sheet can maintain the sheet shape without contracting. When applied to the affected area, the support 4 part can be sandwiched and the inner part of the cell sheet where the support 4 does not exist can be attached and used. Since the shape of the support body 4 is determined by the shape of the sheet-like biological tissue 5, it is not specifically limited. In another embodiment, the support body 4 may not have lost its inner region.

  When the cell sheet in close contact with the support 4 is applied to the container 1 of this embodiment, the edge portion of the cell sheet supported by the support 4 is placed on the lower protrusion 23 (see FIG. 4B). Thereby, the sheet-like living tissue 5 can be separated from the main body lower bottom 21.

  In the present invention, when the main body upper part 3 is fitted to the main body lower part 2, the container 1 includes a gap h formed by the mounting surface 23a and the upper convex part lower surface 33a (see FIG. 4D). By having the gap h, the sheet-like living tissue 5 floats in the space h and is held in a predetermined space. That is, when the main body lower portion 2 and the main body upper portion 3 are combined, the gap h between the lower convex portion 23 and the upper convex portion 33 can hold the sheet-like living tissue 5 in a predetermined space in the accommodation space S. it can. Therefore, the vibration generated when the container 1 is transported is not directly transmitted to the sheet-like living tissue 5, and damage is prevented. The gap h is determined depending on the thickness of the sheet-like living tissue 5 to be accommodated or the thickness of the support 4 to which the sheet-like living tissue is brought into close contact (collectively, “the thickness of the sheet-like living tissue”). Is done. As shown in FIG. 4D, the thickness of the sheet-like living tissue includes the thickness in a state where the sheet-like living tissue 5 is wound around the support 4, and the thickness of the thickest portion of the sheet-like living tissue. Say it. The gap h is preferably the thickness of the sheet-like living tissue +0.1 mm to 5.0 mm, more preferably the thickness of the sheet-like living tissue +0.2 mm to 3.0 mm, and the thickness of the sheet-like living tissue +0.3 mm to More preferably, it is 2.5 mm. The vertical heights of the lower protrusion 23 and the upper protrusion 33 can be determined so that the gap h has a desired distance.

(Constitution example of container (modified example 1))
FIG. 5A shows a modified example (main body lower part 102) of the container according to the embodiment of the present invention. The main body lower part 102 can be used when the sheet-like living tissue 5 is rectangular. In the present embodiment, the main body lower part 102 includes a rectangular main body lower part 121 and a hollow rectangular parallelepiped main part lower part 122 provided on the main part lower part 121. At least a part of the lower body bottom portion 121 is provided with a lower convex portion 123 for placing a part of the peripheral region of the sheet-like living tissue 5 and separating the sheet-like living tissue 5 from the lower body bottom portion 121.

  In the present embodiment, the lower convex portion 123 is disposed in contact with the main body lower bottom portion 121 and the inner wall surface of the main body lower side portion 122. A lower groove 123b is provided between adjacent lower protrusions 123. By providing the lower groove 123b, a jig such as tweezers can be inserted. Thereby, the sheet-like biological tissue 5 can be easily taken out from the main body lower part 102 at the time of use.

  As in the present embodiment, a plurality of lower protrusions 123 may be provided with lower grooves 123b at regular intervals, or may be provided as a hollow cylinder without the lower grooves 123b (not shown). ) As described above, at least one lower groove 123b is preferably provided so that the sheet-like living tissue 5 can be easily taken out during use. The shape and size of the lower groove 123b are not particularly limited as long as the sheet-like living tissue 5 can be taken out with a jig or the like during use.

  The main body upper portion applied to the main body lower portion 102 of the present embodiment can adopt a shape applicable to the main body lower portion 102 (not shown) without departing from the idea of the main body upper portion 3 described above.

(Constitutional example of container (modified example 2))
FIG. 5B shows a modified example (container 201) of the container according to the embodiment of the present invention. The lower body portion 202 includes a hexagonal lower body bottom portion 221 and a hollow hexagonal column lower body side portion 222 provided on the lower body bottom portion 221. At least a part of the lower body bottom portion 221 is provided with a lower convex portion 223 for placing a part of the peripheral region of the sheet-like living tissue 5 and separating the sheet-like living tissue 5 from the lower body bottom portion 221. In the present embodiment, the lower protrusion 223 is provided so as to be parallel to a part of each side of the hexagonal main body lower bottom 221.

  In the present embodiment, the lower protrusion 223 is disposed in contact with the main body lower bottom 221 and the inner wall surface of the main body lower side portion 222. A lower groove 223b is provided between adjacent lower protrusions 223.

  As in the present embodiment, a plurality of lower protrusions 223 may be provided with lower grooves 123b at regular intervals, or may be provided as hollow hexagonal columns without having lower grooves 223b (not shown). ) As described above, it is preferable that at least one lower groove 223b is provided so that the sheet-like living tissue 5 can be easily taken out during use.

  A main body upper portion 203 applied to the main body lower portion 202 of this embodiment includes an air hole 235 and an upper convex portion 233. In this embodiment, the upper convex part 233 has a hollow hexagonal column structure smaller than the inner wall surface of the main body lower side part so as to be fitted to the main body lower side part 22. A part of the upper convex portion 233 has an upper groove 233b structure in which a side surface of the hexagonal column is partially lost. When the main body upper part 203 is fitted to the main body lower part 202, the upper convex part 233 and the lower convex part 223 are spatially alternated. With this structure, when the sheet-like living tissue 5 (not shown) is accommodated, it becomes possible to efficiently communicate between the lower chamber (not shown) and the upper chamber (not shown), and the storage solution in the lower chamber The metabolic gas dissolved and diffused is released by diffusing into the upper chamber 38, and oxygen dissolved in the storage solution in the upper chamber is diffused into the lower chamber.

(Constitution example of container (modification example 3))
FIG. 6 shows a modified example (container 301) of the container in one embodiment of the present invention. The container 301 includes a main body lower portion 302 and a main body upper portion 303, and the basic structure is as described above. The main body lower portion 302 further includes a plurality of leg portions 304. It is preferable that three or more leg portions 304 are provided so as to be kept in parallel when the main body lower portion 302 is placed. In this embodiment, the main body upper surface 331b of the main body upper portion 303 includes a leg insertion portion 305 into which the leg portion 304 can be inserted. By inserting the leg portion 304 into the leg insertion portion 305, the container 301 can be stacked. Thereby, more sheet-like biological tissues can be transported in a small space. The length of the leg part 304 and the depth of the leg insertion part 305 are not limited as long as they do not exceed the thickness of the main body upper ceiling part 331.

  In the present embodiment, the main body upper surface 331b of the main body upper portion 303 is provided with a sealing member attaching portion 306 for attaching the sealing member 6 (not shown). Since the sealing member affixing portion 306 is recessed from the plane of the upper upper surface 331b of the main body, when the sealing member 6 is affixed, unevenness due to the thickness of the sealing member 6 is prevented and is peeled off during transportation. It is prevented. A part of the sealing member attaching portion 306 is provided with a sealing member peeling portion 306a so that the attached sealing member 6 can be easily peeled off.

  In the present embodiment, a main body upper surface 331b of the main body upper portion 303 is provided with a label attaching portion 307 for attaching a label. Since the label sticking portion 307 is recessed from the plane of the upper upper surface 331b of the main body, it is prevented that unevenness due to the thickness of the label is formed when a label (not shown) is stuck, and it is prevented from being peeled off during transportation. Is done. The label sticking unit 307 includes information for identifying the sheet-like living tissue enclosed in the container 301, for example. As the label, a known label such as a barcode, a matrix type two-dimensional code, or an RFID tag can be used. By attaching a label to the container 301, it is possible to prevent mistakes even when a large amount of the container 301 is transported.

(Sheet tissue)
In the present specification, the “sheet-like biological tissue” refers to a biological tissue collected from a living body or a thin-film tissue containing cells. A sheet-like living tissue is a sheet-like living tissue collected from a living body (for example, skin tissue, epithelial tissue, mucosal tissue, fascia, dura mater, periosteum, etc.) or a living tissue obtained by processing a tissue collected from a living body into a sheet shape. There may be. The container 1 of the present invention exhibits an excellent effect particularly in the storage and / or transportation of a fragile sheet-like tissue or a fragile cell sheet collected from a living body.

  In this specification, the “cell sheet” is a single-layer or multi-layer sheet obtained by culturing a cell group containing a plurality of arbitrary cells on a cell culture substrate and peeling the cell group from the cell culture substrate. Cell group. As a method for obtaining a cell sheet, for example, cells are cultured on a stimulus-responsive culture substrate coated with a polymer whose molecular structure is changed by stimulation of temperature, pH, light, etc., and stimulation of temperature, pH, light, etc. By changing the surface conditions of the stimulus-responsive culture substrate and changing the surface conditions, the cells can be detached from the stimulus-responsive culture substrate while maintaining the adhesion state between the cells, or any culture substrate Examples thereof include a method of culturing cells and physically peeling them with tweezers. As a stimulus-responsive culture substrate for obtaining a cell sheet, a temperature-responsive culture substrate having a surface coated with a polymer whose hydration power changes in a temperature range of 0 to 80 ° C. is known. Cells are cultured on a temperature-responsive culture substrate in a temperature range where the polymer hydration power is weak, and then the culture medium is changed to a temperature at which the polymer hydration power is strong. It can be peeled off and collected.

  The temperature-responsive culture substrate used for obtaining the cell sheet is preferably a substrate that changes the hydration power of the surface in a temperature range where cells can be cultured. In general, the temperature range is preferably a temperature for culturing cells, for example, 33 ° C to 40 ° C. The temperature-responsive polymer coated on the culture substrate used for obtaining the cell sheet may be either a homopolymer or a copolymer. Examples of such a polymer include polymers described in JP-A-2-21865.

  A case where poly (N-isopropylacrylamide) is used as a stimulus-responsive polymer, particularly a temperature-responsive polymer will be described as an example (temperature-responsive culture dish). Poly (N-isopropylacrylamide) is known as a polymer having a lower critical solution temperature at 31 ° C. In the free state, dehydration occurs in water at a temperature of 31 ° C. or higher, and polymer chains aggregate and become cloudy. Conversely, at a temperature below 31 ° C., the polymer chain hydrates and becomes dissolved in water. In the present invention, this polymer is coated and fixed on the surface of a substrate such as a petri dish. Therefore, when the temperature is 31 ° C. or higher, the polymer on the surface of the culture substrate is similarly dehydrated. However, since the polymer chain is fixed on the surface of the culture substrate, the surface of the culture substrate is hydrophobic. become. On the other hand, at a temperature lower than 31 ° C., the polymer on the surface of the culture substrate is hydrated, but since the polymer chain is coated on the surface of the culture substrate, the surface of the culture substrate becomes hydrophilic. At this time, the hydrophobic surface is an appropriate surface on which cells can adhere and grow, and the hydrophilic surface is a surface on which cells cannot adhere. Therefore, when the substrate is cooled to less than 31 ° C., the cells are detached from the substrate surface. If the cells are cultured until they are confluent on the entire culture surface, the cell sheet can be recovered by cooling the substrate to less than 31 ° C. The temperature-responsive culture substrate is not limited as long as it has the same effect, and examples thereof include UpCell (registered trademark) marketed by Cellseed Inc. (Tokyo, Japan).

  As the cell sheet stored and / or transported in the present invention, a cell sheet (stacked cell sheet) in which a plurality of cell sheets are stacked may be used. As a method for producing a laminated cell sheet, a cell sheet floating in a culture solution is sucked together with a culture solution by a pipette or the like, and is discharged onto a cell sheet of another culture dish and laminated by a liquid flow. The method of laminating | stacking using a moving jig is mentioned. In addition, a laminated cell sheet can be obtained by a known method.

  In one embodiment, as the support 4 for closely attaching the sheet-like living tissue, for example, polyvinylidene difluoride (PVDF), polyester, polyethylene, polypropylene, silicone resin, polyvinyl alcohol, urethane, cellulose and derivatives thereof, chitin, Chitosan, collagen, gelatin, fibrin gel, metal and the like can be used. The support 4 is used in the form of a film, nonwoven fabric, woven fabric, or mesh. In the embodiment of the present invention, the support 4 is not limited to the above as long as it prevents damage to the sheet-like living tissue without changing the properties thereof.

  The method for bringing the sheet-like living tissue 5 into close contact with the support 4 is not particularly limited, but a method of folding and hooking the sheet-like living tissue 5 that protrudes from the support 4 (for example, FIGS. 4C and D). And a method of adhering to the support 4 using a biocompatible adhesive component (for example, fibrin gel), a method of adhering to the support 4 using an extracellular matrix secreted by the sheet-like living tissue, and the like.

(How to use a container for storage and / or transportation of sheet-like living tissue)
The container 1 of the present invention is used in a method for storing and / or transporting a sheet-like living tissue. In one embodiment, the method comprises:
(1) The sheet-like living tissue 5 is placed on the lower convex portion 23 of the lower body 2 constituting the container,
(2) Apply the upper part 3 of the main body to the lower part 2 of the main body,
(3) The preservation solution 7 is filled from the air hole 35 in the upper part 3 of the main body, the gas is raised along the air slope 37, and the gas is excluded.
(4) Block the air hole 35 with the sealing member 6,
(5) storing and / or transporting the container 1 on which the sheet-like living tissue 5 is placed;
It is a method including.

In one embodiment, the method further comprises:
(6) One or more containers 1 on which the sheet-like living tissue 5 is placed are stored in an airtight secondary container, and stored and / or transported. An airtight secondary container refers to a container in which air does not flow in and out of the secondary container. In this case, the secondary container contains about several hundred mL of air and / or oxygen. The sheet-like living tissue housed in the container of the present invention can be metabolized using this air. The secondary container may contain a CO ₂ absorbent together with the container of the present invention. Moreover, the said secondary container is bundled with the buffer material for absorbing the impact at the time of transport of the container of this invention.

In one embodiment, the method further comprises:
(7) One or more of the secondary containers are stored in a third container having air tightness and heat insulation, and kept at a constant temperature of about 4 ° C to 40 ° C.
including. An airtight tertiary container refers to a container in which air does not flow in and out of the tertiary container. The interior of the tertiary container is maintained at about 1 atmosphere. Moreover, the inside of the tertiary container can be maintained at an arbitrary temperature, for example, a constant temperature of about 4 ° C to 40 ° C. Thereby, it becomes possible to transport the sheet-like living tissue accommodated in the container of the present invention to a place to be used while protecting it from a change in atmospheric pressure, a change in temperature and / or an impact during transportation. That is, according to the present invention, the sheet-like living tissue can be transported to a place where it is used by various transport means while maintaining its activity. For example, it can be transported by any means such as an aircraft, a ship, and a vehicle.

  The preservation solution 7 used in the present invention may be any liquid as long as it does not impair the therapeutic effect of the sheet-like living tissue 5 intended for preservation and transportation. For example, if the sheet-like living tissue 5 is a cell sheet or living tissue, a medium, physiological saline, phosphate buffered saline, or the like suitable for maintaining the cell sheet or living tissue can be used.

  According to the method of the present invention, the sheet-like living tissue 5 can be stored and transported while preventing breakage due to physical vibration and fluid leakage. Further, according to the present invention, the treatment of the sheet-like living tissue 5 that has been conventionally required to be prepared in a limited place (for example, an operating room) immediately before use is performed in a place where the cleanliness is maintained in advance. The sheet-like living tissue 5 can be transported.

  The above-described embodiment is an example of the implementation of the present invention, but the scope thereof is not limited, and various modifications can be made without departing from the scope of the present invention.

  In the above embodiment, the example of the container and the method for accommodating the support to which the sheet-like living tissue is attached has been described. However, the sheet-like living tissue that does not shrink without being attached to the support is also the container of the present invention. And, since it can be stored and / or transported by a method using the same, it is not limited from the scope of the present invention.

1, 201, 301: Container 2, 102, 202, 302: Main body lower portion 21, 121, 221: Main body lower bottom portion 22, 122, 222, 322: Main body lower side portion 22a: Side inner wall surface 22b: Side outer wall surface 22c : O-ring grooves 23, 123, 223, 323: Lower convex portion 23a: Placement surface 23b, 123b, 223b, 323b: Lower groove 24: Key groove 26: O-ring 27, 127, 227, 327: Lower part of main body 28: Lower chamber 3, 203, 303: Main body upper part 31, 331: Main body upper ceiling part 31a: Main body upper inner surface 331b: Main body upper upper surface 32: Main body upper side part 32a: Main body upper side inner wall surface 32b: Main body upper side outer part Wall surface 33, 233, 333: upper convex portion 33a: upper convex portion lower surface 33b, 233b, 333b: upper groove 34: key 35, 235, 335: air hole 6, 236: Main body upper collar portion 37, 237: Air slope 38: Upper chamber 304: Leg portion 305: Leg insertion portion 306: Sealing member attaching portion 306a: Sealing member peeling portion 307: Label attaching portion 4: Support 5: Sheet-like living tissue 6: Sealing member 7: Preservation solution 8: Pipette 9: Bubble S: Storage space h: Gap

Claims (10)

A container for storing and / or transporting a sheet-like living tissue,
A lower part of the main body in which a housing space for accommodating the sheet-like living tissue and the preservation solution is formed by the lower part of the main body and the side of the lower part of the main body
An upper part of the main body which is placed on the upper part of the lower part of the main body to seal the accommodation space and has at least one air hole;
A gas-permeable sealing member for sealing the air hole;
Including
Here, at least a part of the bottom portion of the main body includes a lower convex portion,
Here, an upper convex portion is provided on at least a part of the main body upper inner surface of the main body,
When the lower part of the main body and the upper part of the main body are combined, the gap between the lower convex part and the upper convex part can hold the sheet-like living tissue in a predetermined space in the accommodation space. .   The container according to claim 1, wherein the lower protrusion includes at least one lower groove.   The container according to claim 1, wherein the upper convex portion includes at least one upper groove.   The container according to any one of claims 1 to 3, wherein the upper inner surface of the main body has a slope toward the air hole.   The container according to any one of claims 1 to 4, wherein the sealing member is a liquid-impermeable sealing film.   The container according to any one of claims 1 to 5, wherein the sheet-like living tissue further includes a support.   The container according to claim 6, wherein the support has a shape that supports only a peripheral region of the sheet-like living tissue.   The container according to any one of claims 1 to 7, wherein the upper part of the main body and / or the lower part of the main body includes a collar part. A method for storing and / or transporting a sheet-like living tissue,
(1) A sheet-like living tissue is placed on the lower convex part of the lower part of the main body constituting the container according to any one of claims 1 to 8,
(2) Apply the upper part of the main body to the lower part of the main body,
(3) Fill the storage solution from the air hole to eliminate the gas,
(4) plug the air hole with a sealing member;
(5) storing and / or transporting the container on which the sheet-like living tissue is placed;
Including a method.   The method according to claim 9, wherein the sheet-like biological tissue includes a support.

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