JP7550537B2 - Container for transporting grafts - Google Patents

Description

The present invention relates to a container for transporting a graft.

In recent years, new regenerative medicine has been developed as a solution to the treatment of severe heart failure. One example is a method in which a sheet-shaped cell culture made using a temperature-responsive culture dish using tissue engineering is applied to the surface of the heart in severe myocardial infarction. The method using the sheet-shaped cell culture makes it possible to safely transplant a large amount of cells over a wide area, and is particularly useful for the treatment of myocardial infarction (including chronic heart failure associated with myocardial infarction), dilated cardiomyopathy, ischemic cardiomyopathy, and heart diseases (e.g., heart failure, especially chronic heart failure) accompanied by systolic dysfunction (e.g., left ventricular systolic dysfunction), etc.

For clinical application of such sheet-shaped cell cultures, 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 or other location where the transplant will take place. However, sheet-shaped cell cultures have low absolute physical strength and can become wrinkled, torn or damaged by vibrations that occur when transporting the container, so this transport process requires advanced techniques and meticulous care.

To meet these needs, various methods and containers have been developed. For example, the storage and transport container for membranous tissue described in Patent Document 1 below fills the storage section with a preservation liquid to the extent that no gas layer is formed, so that the preservation liquid does not ripple or flow. As a result, vibrations are not transmitted to the membranous tissue, and damage to the membranous tissue can be prevented.

The following Patent Document 2 describes a container for holding a sheet-like graft. Such a container has a sealable bag body that contains a sheet-like graft together with a liquid, and the bag body is provided with a graft removal outlet and an opening for inserting the sheet-like graft, and the graft removal outlet can be sealed with a lid via a peelable weak seal portion.

JP 2012-130311 A JP 2013-215168 A

As mentioned above, containers have been developed to safely transport cell sheets, and the inside of the container is made liquid-tight to prevent the movement of air bubbles inside.

However, advanced technology is required to completely remove air bubbles from inside a container like the one described above and achieve a liquid-tight state. Furthermore, if a bag that is prone to deformation as described above is used, it can easily deform and damage the cell sheet inside if it is dropped or subjected to pressure during transport.

The present invention was made with these points in mind, and aims to provide a container that can achieve a liquid-tight state efficiently with simple operations and can transport grafts stably.

That is, the present invention relates to the following.
[1] A container for transporting a graft, comprising: a bag body capable of containing the graft and being liquid-tightly sealed; and a main body portion for containing the bag body, the main body portion having a fixing portion for fixing the bag body, the container being configured so that the bag body is in an expanded state when the bag body is fixed to the fixing portion.
[2] The container described in [1], wherein the fixing portion is configured so that the bag body can be fixed in a floating state within the main body portion.
[3] A container according to [1] or [2], wherein the fixing portion is spaced apart from the inner surface of the main body portion.
[4] A container according to any one of [1] to [3], wherein the bag body does not come into contact with the main body portion by expanding the bag body.

[5] A container as described in [4], wherein the fixing portion is configured to be able to fix the bag body in an expanded state at a position spaced from the inner surface of the main body portion.
[6] The container according to any one of [1] to [5], wherein the bag body has an exhaust port for exhausting air inside the bag body.
[7] A container described in any one of [1] to [6], wherein the main body includes a base portion having a fixing portion and a cover portion for covering the base portion, and the base portion is configured so that the bag body can be fixed in a vertically placed position.
[8] The container according to any one of [1] to [7], wherein the graft is a sheet-shaped cell culture.
[9] The container according to [8], wherein the sheet-shaped cell culture is a laminate.

The bag body of the present invention has a simple mechanism and can efficiently achieve a liquid-tight state with simple operations, and therefore has great advantages in terms of workability and manufacturing costs. Furthermore, because the inside of the bag body can be made completely liquid-tight, air bubbles (gas) do not enter the bag body, and air bubbles do not move inside the bag body due to shaking of the container during transport, etc., and do not damage the graft. In particular, when the graft is a laminate of sheet-shaped cell cultures, air bubbles do not move and cause the laminate to shift or become damaged. Therefore, the shape of the graft in the liquid can be maintained and deformation prevented, allowing long-term storage.

The main body of the present invention contains the bag body, and the fixing part provided inside reliably fixes the bag body, and when the bag body is fixed to the fixing part, the bag body can be in an expanded state, so that the bag body does not deform and the shape of the graft inside the bag body can be maintained. Furthermore, the robustness of the main body part allows the bag body to be protected from external impact, and the bag body can be fixed in a floating state within the main body part, so that the bag body and the fixing part can form an impact-resistant damper. Furthermore, the base part of the main body part allows the bag body to be fixed in an upright position, so that the bag body can be conveniently housed in the bag body, sealed, and liquid-tightly sealed.

FIG. 1 is a conceptual diagram of a main body 1 according to a first embodiment of the present invention. FIG. 2 is a conceptual diagram of the bag body 2 housed in the main body 1 shown in FIG. FIG. 3 is a conceptual diagram showing the bag body 2 in a deployed state within the main body 1. As shown in FIG.

In the present invention, a graft refers to a fragile object derived from a living organism that has relatively low physical strength. The graft includes cultured cells (e.g., cell cultures, etc.) and harvested cells. The graft may further include products produced by cells. In addition to cells and/or cell products, the graft may also include materials (filling materials and support materials) for filling and/or supporting a specific part of the organism (e.g., an affected area, etc.). The graft may have various shapes such as a sheet, a membrane, a block, a column, etc. The graft is used for transplantation into a living organism. Examples of grafts include three-dimensional cell tissues (organoids, spheroids, etc.) and two-dimensional cell tissues (sheet-shaped cell cultures, etc.).

In the present invention, a sheet-shaped cell culture refers to a sheet-shaped structure in which cells are connected to each other. The cells may be connected to each other directly (including through cellular elements such as adhesion molecules) and/or through an intervening substance. The intervening substance is not particularly limited as long as it can at least physically (mechanically) connect the cells to each other, and examples thereof include extracellular matrices. The intervening substance is preferably derived from cells, particularly from the cells that constitute the sheet-shaped 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 (multilayer). The sheet-shaped cell culture may have a three-dimensional structure in which the cells do not show a clear layer structure and have a thickness greater than the thickness of one cell. For example, in a vertical cross section of the sheet-shaped cell culture, the cells may be present in a state in which they are arranged unevenly (for example, in a mosaic shape) without being uniformly aligned in the horizontal direction. The sheet-shaped cell culture may exist as a single (one) independently formed sheet-shaped cell culture, or as a laminate formed by stacking two or more independent single (one) sheet-shaped cell cultures. The laminate may be, for example, a laminate formed by stacking two layers (two sheets), three layers (three sheets), four layers (four sheets), five layers (five sheets), or six layers (six sheets) of sheet-shaped cell cultures.

The sheet-shaped cell culture of the present invention is composed of any cells capable of forming the above-mentioned structure. Examples of such cells include, but are not limited to, adhesive cells (adherent cells). Adherent cells include, for example, adhesive somatic cells (e.g., cardiomyocytes, fibroblasts, epithelial cells, endothelial cells, hepatic cells, 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 be differentiated from stem cells, particularly iPS cells. Non-limiting examples of cells that can form a sheet-shaped cell culture include myoblasts (e.g., skeletal myoblasts, etc.), mesenchymal stem cells (e.g., derived from bone marrow, adipose tissue, peripheral blood, skin, hair roots, muscle tissue, endometrium, placenta, umbilical cord blood, etc.), 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, etc.), endothelial cells (e.g., vascular endothelial cells, etc.), hepatic cells (e.g., hepatic parenchymal cells, etc.), pancreatic cells (e.g., pancreatic islet cells, etc.), kidney cells, adrenal cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, etc. In the present specification, those that form a monolayer cell culture, such as myoblasts or cardiomyocytes, are preferred, and particularly preferred are cardiomyocytes derived from skeletal myoblasts or iPS cells.

The cells may be derived from any organism that can be treated with cell culture. Such organisms include, but are not limited to, humans, non-human primates, dogs, cats, pigs, horses, goats, sheep, and the like. In addition, only one type of cell may be used to form the sheet-shaped cell culture, but two or more types of cells can also be used. In a preferred embodiment of the present invention, when there are two or more types of cells forming the cell culture, the proportion (purity) of the most abundant cells is 65% or more, preferably 70% or more, and more preferably 75% or more, for example, in the case of skeletal myoblasts, at the end of the cell culture production.

The sheet-shaped cell culture of the present invention may be a sheet-shaped cultured tissue obtained by seeding cells on a scaffold (a base for cell culture) and culturing the cells, but preferably consists only of substances derived from the cells that constitute the cell culture and does not contain any other substances.
The sheet-shaped cell culture may be produced by any known method.

In one embodiment of the present invention, the sheet-shaped cell culture is a sheet-shaped skeletal myoblast culture. This is because sheet-shaped skeletal myoblast cultures are so fragile that they break under their own weight when part of them is grasped, and therefore not only have they not been able to be transported alone in the past, but once they are folded over, it is extremely difficult to return them to their original shape, so maintaining the sheet shape in liquid is of great significance.

In the present invention, the bag body is not particularly limited as long as it can contain a graft, liquid, etc. inside and does not leak liquid, and any material including commercially available containers can be used. As a material for the bag body, a soft resin material with excellent flexibility and suppleness is preferable. This ensures the flexibility of the bag body and facilitates the injection of liquid and the insertion and removal of grafts. Such resin materials are not particularly limited, but examples include polyolefins such as polyethylene, polypropylene, polybutadiene, and ethylene-vinyl acetate copolymers, polyesters such as polyethylene terephthalate and polybutylene terephthalate, soft polyvinyl chloride, polyvinylidene chloride, silicone, polyurethane, styrene-butadiene copolymers, polyamide elastomers, polyester elastomers, and any combinations of these (blend resins, polymer alloys, laminates, etc.). In terms of sealability, heat resistance, water resistance, flexibility, and processability, polypropylene-based soft resins are particularly preferable. In addition, from the viewpoints of visibility of the amount of liquid in the bag body and confirmation of the position of the graft, a resin material with transparency is preferable.

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 other liquid. The liquid may be a fluid that has overall fluidity, and may contain solid substances such as cell scaffolds, or other non-liquid components such as air bubbles.

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 derived from a living organism, the components constituting the liquid in the container are preferably biocompatible from the viewpoint of biological stability and long-term storage potential, that is, components that do not cause undesirable effects such as inflammatory reactions, immune reactions, or toxic reactions in living tissues or cells, or at least have small 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, RPMI1640, MC100, etc.), and the like. DB, L15, SkBM, RITC80-7, IMDM, etc.), sugar solution (sucrose solution, Ficoll-paque (registered trademark) PLUS, etc.), seawater, serum-containing solution, 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.

When the graft is a sheet-shaped cell culture, the components constituting the liquid in the container are preferably capable of stably storing the cells, contain the minimum amount of oxygen and nutrients necessary for cell survival, and do not destroy the cells due to osmotic pressure, etc. 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.

In the present invention, the main body is not particularly limited as long as it can accommodate the bag body inside and is robust and impact resistant, and any material including commercially available containers can be used. Materials for the main body 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, metals (e.g., iron, stainless steel, aluminum, copper, brass), etc.

In the present invention, the shapes of the bag body and the main body are not particularly limited, so long as the bag body can be fixed within the main body and the bag body is in an expanded state as a result of such fixing. For example, if the bag body is rectangular, it is preferable that the main body be rectangular in shape. In addition, the bag body and/or the main body may be made of a light-transmitting material, so that the state of the graft contained in the bag and the presence or absence of air bubbles in the liquid can be confirmed.

First Embodiment
One aspect of the present invention relates to a container for transporting a graft, the container including a bag body capable of containing the graft and being liquid-tightly sealed, and a main body portion for containing the bag body, the main body portion having a fixing portion for fixing the bag body, and configured so that the bag body is in an expanded state when the bag body is fixed to the fixing portion.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings.
First, a first embodiment of the present invention will be described.
Fig. 1 is a conceptual diagram of a main body 1 according to a first embodiment of the present invention, and Fig. 2 is a conceptual diagram of a bag body 2 housed in the main body 1 shown in Fig. 1. In each drawing in the present application, the size of each member is appropriately exaggerated for ease of explanation, and the illustrated members do not show their actual size.

As shown in FIG. 1, the main body 1 is a vertically long rectangular cube in one embodiment, and is designed to suitably accommodate the vertically long bag body 2 inside. The main body 1 has a plurality of fixing parts 12 inside, which are columnar and spaced apart from the inner surface of the main body 1. In one embodiment, the main body 1 can include a base 11 having the fixing parts 12, and a cover 14 for covering the base 11. The base 11 is configured so that the bag body 2 can be fixed in a vertically placed state.

That is, the base 11 is configured such that two plate-like members are connected perpendicular to each other, one surface is stably placed on a workbench, and the fixing part 12 is provided on another surface that is erected perpendicular to the first surface, so that the bag body 2 can be fixed in an upright position. The cover 14 is a vertically long rectangular cube, with an opening 15 on one side for receiving the base 11, and the base 11 to which the bag body 2 is fixed can be slid in through the opening 15 to house the bag body 2 inside, and the cover 14 can be sealed by using the above-mentioned one surface as a lid.

As shown in FIG. 2, the bag body 2 is a rectangular cube in one embodiment, and is dimensioned so that the graft G can be suitably accommodated in the expanded state inside the bag body 2. The peripheral portion of the bag body 2 is provided with a plurality of holes 21 through which the fixing parts 12 can be inserted. The number of holes 21 corresponds to the number of fixing parts 12, and the bag body 2 is positioned so that the bag body 2 is expanded when the bag body 2 is fixed to the fixing parts 12, i.e., the bag body 2 is pulled in the left-right and up-down directions. The bag body 2 has an opening 23 at one end for inserting the graft G, and an exhaust port 22 at the other end for exhausting the air inside the bag body 2.

The procedure for storing the graft G in the bag body 2 will be briefly described. This procedure will be described assuming that it is performed in a clean room. When storing the graft G in the bag body 2, first, the bag body 2 is fixed vertically on the base part 11 so that the opening 23 is on top, and the discharge port 22 is sealed at the sealing position S by heat sealing or the like (Fig. 2a). Next, the graft G and liquid are inserted from the opening 23 of the bag body 2 (Fig. 2b). Since the bag body 2 is fixed vertically, the liquid can be poured without leaking out of the bag body 2, and the opening 23 can be suitably sealed at the sealing position S by heat sealing or the like (Fig. 2c). By providing an easy peel at the sealing position S of the bag body 2, it is possible to easily open and close the seal.

When the opening 23 is sealed, air may enter the bag body 2, and this air must be removed. In this case, the bag body 2 is turned upside down to move the air toward the discharge port 22 (Fig. 2d). At this time, the other side of the bag body 2, where the discharge port 22 is located, may be tapered so that the buoyant air can be configured to move favorably toward the discharge port 22 along the tapered portion. Then, a syringe or the like is connected to the discharge port 22 to suck out the air inside, and the bag body 2 is sealed at the sealing position S, allowing the graft G to be contained within the bag body 2 and sealed liquid-tightly.

In one embodiment, the bag body 2 can be provided with a columnar fixing part 12 having a recess 13 in its center, and the bag body 2 can be fixed in a floating state within the main body part 1 by engaging the hole 21 of the bag body 2 with the recess 13. That is, the length of the fixing part 12 and the position of the recess 13 are designed to be of sufficient length and position so that both sides of the bag body 2 do not come into contact with the inner surface of the main body part 1 when the bag body 2 is fixed to the recess 13. The fixing part 12 is provided away from the inner surface of the main body part 1 so that the peripheral part surrounding the bag body 2 does not come into contact with the inner surface of the main body part 1. Then, the base part 11 can be slid into the cover part 14 without removing the bag body 2 from the base part 11 where the work of storing it has been performed, thereby sealing the bag body 2 within the container 1 and protecting the bag body 2 within the highly robust container 1.

By transporting the container 1 configured as above, even if the bag body 2 is easily deformed, the graft G inside is not damaged even if it is dropped or pressure is applied during transport. When removing the bag body 2 at the destination, the base part 11 can be slid out from the cover part 14 to easily remove it. If the bag body 2 is heat sealed, the graft G contained therein can be easily removed by cutting it with scissors at the sealing position S at the destination, or if it is easy peel, by peeling it. Since the bag body 2 is required to be sterilized, for example, the bag body 2 may be transported in a state in which it is placed in a second bag body 2' (not shown). The peripheral part of the second bag body 2' may also be provided with a similar number of holes 21', and the holes 21 of the bag body 2 and the holes 21' of the second bag body 2' may be inserted into the fixing part 12 to transport the bag body 2 while ensuring its sterility.

While the container 1 according to the present invention has been described above based on the first embodiment, the present invention is not limited to this and can have various modifications.
In the present invention, the fixing part 12 is not particularly limited as long as it is configured so that the bag body 2 can be fixed in a stretched state at a position separated from the inner surface of the main body part 1, that is, so that the bag body 2 is fixed at a position where it does not come into contact with the main body part 1, as shown in Fig. 3. A part of the bag body 2 may come into slight contact with the main body part 1, and may come into contact to such an extent that the weight of the bag body 2 is hardly applied to the main body part 1. For example, it may be configured so that the gravity from the bag body 2 to the main body part 1 is smaller than the horizontal stretching force of the bag body 2.
For example, the fixing part may be configured as a column having a screw hole on its upper surface, and the bag body may be fixed to the same position as the recess 13 of the fixing part 12 in FIG. 1 by screwing the bag body into the screw hole. In this case, the multiple holes 21 may be present or absent. Also, for example, the fixing part 12 does not need to be configured as a column as described above, and may be in the form of a hook that can hook the multiple holes 21. Also, the bag body 2 can be fixed to the main body 1 in various ways. For example, the multiple holes 21 may be omitted, and the fixing part 12 may be configured as a double clip or a paper clip, and the peripheral portion of the bag body 2 may be clamped to fix the bag body in an expanded state.
In the present invention, each component can be replaced with any component capable of exerting a similar function, or any component can be added.

As described above, the bag body of the present invention has a simple mechanism and can efficiently achieve a liquid-tight state with simple operations, and therefore has great advantages in terms of workability and manufacturing costs. Furthermore, because the inside of the bag body can be made completely liquid-tight, air bubbles (gas) do not enter the bag body, and air bubbles do not move inside the bag body due to shaking of the container during transport, etc., and do not damage the graft. In particular, when the graft is a laminate of sheet-shaped cell cultures, air bubbles do not move and cause the laminate to shift or become damaged. Therefore, the shape of the graft in the liquid can be maintained and deformation prevented, allowing long-term storage.

The main body of the present invention contains the bag body, and the fixing part provided inside reliably fixes the bag body, and when the bag body is fixed to the fixing part, the bag body can be in an expanded state, so that the bag body does not deform and the shape of the graft inside the bag body can be maintained. Furthermore, the robustness of the main body part allows the bag body to be protected from external impact, and the bag body can be fixed in a floating state within the main body part, so that the bag body and the fixing part can form an impact-resistant damper. Furthermore, the base part of the main body part allows the bag body to be fixed in an upright position, so that the bag body can be conveniently housed in the bag body, sealed, and liquid-tightly sealed.

REFERENCE SIGNS LIST 1 Main body 11 Base 12 Fixing portion 13 Recess 14 Cover 15 Opening 2 Bag body 21 Hole 22 Discharge port 23 Opening G Graft S Sealing position

Claims (11)

A container for transporting a graft, comprising: a bag body capable of containing the graft and being liquid-tightly sealed; and a main body portion for containing the bag body, the main body portion having a fixing portion for fixing the bag body, the bag body being in an expanded state when the bag body is fixed to the fixing portion , and the fixing portion being configured so that the bag body can be fixed in a floating state within the main body portion . 2. The container according to claim 1, wherein the fixing portion is positioned so that the bag body is pulled and expanded when the bag body is fixed to the fixing portion. 3. The container according to claim 1 or 2, wherein the fixing portion has a recess and/or a screw hole, and is configured so that the bag body can be fixed in a floating state within the main body portion by fixing the bag body to the recess and/or the screw hole. The container according to any one of claims 1 to 3, wherein the bag body has a plurality of holes, and is configured so that the plurality of holes can be fixed to a plurality of fixing parts. The container according to any one of claims 1 to 4, wherein the fixing portion is provided spaced apart from an inner surface of the main body portion. The container according to any one of claims 1 to 5 , wherein the bag body is not in contact with the main body by being expanded. 7. The container according to claim 6 , wherein the fixing portion is configured to be capable of fixing the bag body in a stretched state at a position spaced from the inner surface of the main body. The container according to any one of claims 1 to 7 , wherein the bag body has an exhaust port for exhausting air inside the bag body. The container according to any one of claims 1 to 8, wherein the main body includes a base having a fixing portion and a cover for covering the base, and the base is configured so that the bag body can be fixed in a vertically placed state. The container according to any one of claims 1 to 9 , wherein the graft is a sheet-like cell culture. The container according to claim 10 , wherein the sheet-shaped cell culture is a laminate.