WO2020013174A1

WO2020013174A1 - Device for transporting grafts

Info

Publication number: WO2020013174A1
Application number: PCT/JP2019/027128
Authority: WO
Inventors: 鮫島　正; 健太水本
Original assignee: テルモ株式会社
Priority date: 2018-07-10
Filing date: 2019-07-09
Publication date: 2020-01-16
Also published as: JPWO2020013174A1; WO2020012452A1; JP7319976B2

Abstract

The purpose of the present invention is to provide a device that makes it possible to efficiently achieve liquid-tightness by means of a simple operation and to easily collect liquid from inside a container for use as a test sample. The present invention relates to a device that can be detachably installed on a container that is for storing grafts. The device includes a lid member that is for sealing the container. The lid member has a top plate part that covers an opening part of the container. The top plate part has a dome-shaped protruding part, a liquid-discharge opening part, and a liquid-injection opening part. The liquid-discharge opening part is arranged at an apex part of the protruding part.

Description

Device for transferring an implant

The present invention relates to a device for transferring an implant.

In recent years, the development of new regenerative medicine as a solution for the treatment of severe heart failure has been promoted. As an example, a method of applying a sheet-shaped cell culture prepared using a temperature-responsive culture dish to which tissue engineering is applied in severe myocardial infarction or the like to the heart surface has been attempted. The method using a sheet-shaped cell culture allows a large amount of cells to be transplanted safely over a wide area, such as myocardial infarction (including chronic heart failure associated with myocardial infarction), dilated cardiomyopathy, ischemic cardiomyopathy It is particularly useful for the treatment of heart diseases associated with systolic dysfunction (eg, left ventricular systolic dysfunction) (eg, heart failure, especially chronic heart failure).

臨床 To apply such a sheet-shaped cell culture to clinical application, for example, it is necessary to store the prepared sheet-shaped cell culture together with a preservation solution in a container and transfer it to an intensive care unit where transplantation is performed. However, sheet cell cultures have low absolute physical strength, and wrinkles, tears, breakage, etc. occur due to vibrations generated when transferring containers, etc. You need to be very careful.

種々 Various methods and containers have been developed to meet such needs. For example, the container for storing and transporting a membrane-like tissue described in Patent Document 1 below fills the storage portion with a storage solution to such an extent that a gas layer is not formed, thereby causing the storage solution to undulate or flow. As a result, vibration is not transmitted to the film-like tissue, and damage to the film-like tissue can be prevented.

包装 Patent Document 2 below describes a package for transporting a cultured cell sheet. Such a package eliminates air with some liquid medium when sealing with a lid, so that even if the cultured cell sheet package shakes during transportation, bubbles do not move in the liquid medium, It is designed to prevent displacement and loss of the cultured cell sheet.

JP 2012-130311 A JP 2009-89715 A

容器 Containers have been developed to safely transport fragile cell sheets as described above. Further, such a container is devised so that the inside of the container is liquid-tight so that the movement of air bubbles does not occur.

However, advanced technology is required to completely remove air bubbles from the container and achieve a liquid-tight state. In addition, since a strong force is required to remove the lid from the container in a liquid-tight state, the liquid may wave when the lid is attached or detached, and the cell sheet may be damaged.

The present invention has been made in view of such points, and an object of the present invention is to provide a device that can efficiently achieve a liquid-tight state with a simple operation.

That is, the present invention relates to the following.
[1] A device used detachably mounted on a container for accommodating an implant, wherein a lid member for sealing the container and a first space are formed between the lid member and the container. The device, wherein a second space is formed between the lid member and the inner surface of the container.
[2] The device according to [1], further including a seal member interposed between the container and the lid member.
[3] The device according to [1] or [2], further including a protection member interposed between the annular member and the lid member to protect the first space.

[4] The device according to [3], wherein the protection member has flexibility.
[5] The device according to [1] or [3], wherein the protective member and / or the lid member are subjected to a water-repellent treatment.
[6] The device according to any one of [1] to [5], wherein the implant is a sheet-shaped cell culture.
[7] The device according to [6], wherein the sheet-shaped cell culture is a laminate.
[8] A holding device including a cover member and a base member configured to be able to press and hold the device according to any one of [1] to [7] from above and below.

According to the device of the present invention, since a liquid-tight state can be efficiently achieved by a simple mechanism and a simple operation, there are great advantages in terms of workability and manufacturing cost. And since the inside of a container can be made into a completely liquid-tight state, air bubbles (gas) do not enter in a container and bubbles do not move in a container due to shaking of the container during transfer, and the fragile object is not damaged. In particular, when the fragile material is a laminate of sheet-shaped cell cultures, bubbles do not move, and the laminate does not shift or lose. Therefore, it can be stored for a long time while maintaining the shape of the fragile object in the liquid to prevent deformation.

Since the device of the present invention can easily release the liquid-tight state, it is possible to suppress the occurrence of vibration of the container and the flow of the liquid due to, for example, forcible removal of the lid member of the worker, and to reduce fragile objects. No damage. In addition, since work can be performed without contaminating the surroundings, cleanliness is strictly controlled, such as in a bio-clean room used for producing sheet-shaped cell cultures and in intensive care rooms where sheet-shaped cell cultures are used. Suitable for use in controlled locations.

FIG. 1 is a conceptual diagram of a device 1 according to the first embodiment. FIG. 2 is a cross-sectional view of the device 1 shown in FIG. FIG. 3 is a perspective view of the holding device 7 according to the second embodiment. FIG. 4 is a perspective view of a holding tool 7A according to the third embodiment. FIG. 5 is a perspective view of a holding tool 7B according to the fourth embodiment.

脆弱 A fragile object in the present invention refers to an object that has low physical strength and can be broken, broken, deformed, or the like due to shaking of a liquid. Examples of the shape of such an object include an object having a thin portion, an object having a band shape, an object having a sheet shape, and the like. Examples of the object having such a sheet shape include, but are not particularly limited to, a sheet-like structure, for example, a sheet-like cell culture, a flat membrane-like membrane tissue made of a biological material, plastic, paper, woven fabric, and nonwoven fabric. And films of various materials such as metals, polymers, and lipids. Among them, those which are hardly decomposable in liquid and those which are hardly disintegratable in liquid are preferred. The sheet-like structure may be polygonal or circular, and may or may not be uniform in width, thickness, diameter, and the like. The sheet-like structure in the present invention may be used alone in a single layer state, or may be used in a laminated state in which two or more sheets are stacked. In the latter case, each layer of the laminate may be connected to each other, or may not be connected.If connected, all the overlapping portions may be connected or partially connected. Is also good. Further, in the present invention, the fragility is, for example, evaluation of the tensile properties in a conventional tensile tester (for example, those described in JIS K 7161 etc.) in which the object is fixed to the gripper outside the liquid, Difficult or practically impossible due to vulnerability. As such a fragile object, for example, an object having a small value of each of the tensile properties and difficult to accurately measure with a conventional tensile tester can be cited. Examples of such brittle materials include those that exhibit a breaking load of less than 10 N (Newton), less than 5 N, less than 2 N, less than 1 N, less than 0.5 N, less than 0.1 N, and less than 0.05 N in a tensile test. . In addition, since the measurement limit of the conventional tensile test is generally about 1 N as a breaking load, in one embodiment of the present invention, a material showing a breaking load lower than this (for example, less than 0.5 N) is preferable as a brittle substance. .

In the present invention, the term “graft” refers to a fragile substance derived from a living body having relatively low physical strength. Grafts include cultured cells (eg, cell cultures) and harvested cells. Implants may also contain products produced by the cells. The graft may contain, in addition to the cells and / or the products of the cells, a material (supplementary material or support material) for supplementing and / or supporting a predetermined part (for example, an affected part) of the living body. The implant can take various shapes such as a sheet, a film, a block, and a column. The graft is used for transplantation into a living body or the like. Examples of the transplant include a three-dimensional cell tissue (organoid, spheroid, etc.) and a two-dimensional cell tissue (sheet-shaped cell culture, etc.).
In the present invention, the sheet-shaped cell culture refers to a cell in which cells are connected to each other to form a sheet. The cells may be connected to each other directly (including via a cell element such as an adhesion molecule) and / or via an intermediary substance. The intervening substance is not particularly limited as long as it is a substance capable of at least physically (mechanically) connecting cells to each other, and examples thereof include an extracellular matrix. The intervening substance is preferably derived from cells, particularly from cells constituting a sheet-shaped cell culture. Cells are at least physically (mechanically) linked, but may be further functionally linked, for example, chemically or electrically. The sheet-shaped cell culture may be composed of one cell layer (single layer) or composed of two or more cell layers (multilayer body). Further, the sheet-shaped cell culture may have a three-dimensional structure having a thickness exceeding the thickness of one cell without the cells showing a clear layer structure. For example, in the vertical cross section of the sheet-shaped cell culture, the cells may exist in a non-uniform (eg, mosaic) state without being uniformly arranged in the horizontal direction. The sheet cell culture may exist as a single, independently formed sheet cell culture, or an independent single sheet cell culture may be present. It may exist as a laminate formed by laminating two or more layers. The layered product is, for example, a layered structure obtained by laminating two (2), three (3), four (4), five (5), or six (6) sheet-shaped cell cultures. It can be a body.

シート 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, adherent cells (adherent cells). Adherent cells include, for example, adherent somatic cells (eg, 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 (eg, tissue stem cells such as myoblasts, heart stem cells, embryonic stem cells, pluripotent stem cells such as iPS (induced pluripotent stem) cells, mesenchymal stem cells, etc.) Including. The somatic cells may be those differentiated from stem cells, especially iPS cells. Non-limiting examples of cells that can form sheet-shaped cell cultures include, for example, myoblasts (eg, skeletal myoblasts), mesenchymal stem cells (eg, bone marrow, adipose tissue, peripheral blood, skin, hair roots) , Muscle tissue, endometrium, placenta, cord blood, etc.), cardiomyocytes, fibroblasts, cardiac stem cells, embryonic stem cells, iPS cells, synovial cells, chondrocytes, epithelial cells (eg, oral mucosal epithelium) Cells, retinal pigment epithelial cells, nasal mucosal epithelial cells, etc.), endothelial cells (eg, vascular endothelial cells, etc.), hepatocytes (eg, hepatic parenchymal cells, etc.), pancreatic cells (eg, pancreatic islet cells, etc.), kidney cells, adrenal glands Cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells and the like. In the present specification, those forming a monolayer cell culture, such as myoblasts or cardiomyocytes, are preferred, and skeletal myoblasts or iPS cell-derived cardiomyocytes are particularly preferred.

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

The sheet-shaped cell culture in the present invention may be a sheet-shaped cultured tissue obtained by disseminating and culturing cells in a scaffold (a scaffold for cell culture), but preferably constitutes a cell culture. Consists of only cell-derived substances and contains no other substances.
The sheet cell culture may be produced by any known technique.

において In one embodiment of the present invention, the sheet-shaped cell culture is a sheet-shaped skeletal myoblast cell culture. This is because the sheet-shaped skeletal myoblast culture is so fragile that it breaks under its own weight when a part of it is grasped, so it cannot be transported alone by itself, nor can it return to its original shape once folded. This is because it is extremely difficult to maintain the shape of the sheet in the liquid.

In the present invention, the container is not particularly limited as long as it can store a fragile substance, a liquid, or the like and does not leak the liquid, and any container including a commercially available container can be used. Examples of the material of the container include polyethylene, polypropylene, Teflon (registered trademark), polyethylene terephthalate, polymethyl methacrylate,

nylon

6,6, polyvinyl alcohol, cellulose, silicon, polystyrene, glass, polyacrylamide, polydimethylacrylamide, and metal ( Examples thereof include, but are not limited to, iron, stainless steel, aluminum, copper, and brass). In addition, the container preferably has at least one flat bottom surface for maintaining the shape of the fragile object, and examples thereof include, but are not limited to, a petri dish, a cell culture dish, a cell culture bottle, and the like. The area of the flat bottom surface is not particularly limited, but is typically 9.1 to 78.5 cm ² , 1.13 to 78.5 cm ² , preferably 12.6 to 78.5 cm ² , and more preferably 9 to 98.5 cm ² . .1 to 60.8 cm ² .

In the present invention, the liquid in the container is composed of at least one kind of component, and the component is not particularly limited. For example, it is composed of liquid such as water, aqueous solution, non-aqueous solution, suspension, and emulsion.
The liquid or liquid in the present specification may be a fluid having fluidity as a whole, and may include other non-liquid components such as a solid substance such as a cell scaffold and bubbles.

成分 The components constituting the liquid in the container are not particularly limited as long as they have little effect on the fragile substance. When the fragile substance is a membrane made of a bio-derived material, the components constituting the liquid in the container are biocompatible from the viewpoint of biological stability and long-term storability, i.e. On the other hand, those which do not cause unwanted effects such as inflammatory response, immune response, and poisoning response or at least have small effects are preferable. For example, water, saline, physiological buffer (eg, HBSS, PBS, EBSS, Hepes) , Sodium bicarbonate, etc.), medium (eg, DMEM, MEM, F12, DMEM / F12, DME, RPMI1640, MCDB, L15, SkBM, RITC80-7, IMDM, etc.), sugar solution (sucrose solution, Ficoll-paque (registered)) Trademark) PLUS), seawater, serum-containing solution, renographin (registered trademark) solution, metrizamide solution, meg Min solution, glycerin, ethylene glycol, ammonia, benzene, toluene, acetone, ethyl alcohol, benzol, oil, mineral oil, animal fat, vegetable oil, olive oil, colloid solution, liquid paraffin, turpentine oil, linseed oil, castor oil, etc. Can be

When the fragile material is a sheet-shaped cell culture, the components constituting the liquid in the container can stably store the cells, contain the minimum amount of oxygen and nutrients necessary for cell survival, and Preferred are those that are not destroyed by osmotic pressure or the like. For example, physiological saline, physiological buffers (eg, HBSS, PBS, EBSS, Hepes, sodium bicarbonate, etc.), culture media (eg, DMEM, MEM, F12, DMEM / F12, Examples include, but are not limited to, DME, RPMI 1640, MCDB, L15, SkBM, RITC80-7, IMDM, and the like, and sugar solutions (sucrose solution, Ficoll-paque @ PLUS (registered trademark), and the like).

The amount of liquid in the container is such that the fragile object can be held in a state where the lid member is attached to the container, and the volume of liquid formed between the bottom of the container and the top of the lid member is reduced. The height is not particularly limited as long as it does not swing (do not swing) in the interior. That is, when a space is formed between the liquid surface and the lid member, the liquid (liquid surface) swings, and the fragile object easily swings in the liquid, so that there is no space between the liquid surface and the lid member. It is preferable to adjust the liquid volume. In one embodiment of the present invention, the diameter of the sheet cell culture is about 35 to 55 mm and the area is 6 cm ² or more, or 10 cm ² or more. The liquid volume can be, for example, 1.0 mm to 20.0 mm regardless of the diameter of the sheet-shaped cell culture.

において In the present invention, the lid member is not particularly limited as long as it can seal the container. Examples of the material of the lid member include polyethylene, polypropylene, Teflon (registered trademark), polyethylene terephthalate, polymethyl methacrylate,

nylon

6,6, polyvinyl alcohol, cellulose, silicon, polystyrene, glass, polyacrylamide, polydimethylacrylamide, and metal. (Eg, iron, stainless steel, aluminum, copper, brass) and the like, but are not limited thereto.

において In the present invention, the shapes of the lid member and the container are not particularly limited as long as the lid member and the container can be engaged with each other and a closed space can be formed by such engagement. For example, when the container is a general-purpose petri dish, the shape of the lid member is preferably circular. The lid member and / or the container may be made of a light-transmitting material so that the state of the fragile object contained in the container and the presence or absence of bubbles in the liquid may be confirmed.

において In the present invention, the fragile object is held in the liquid in the container containing the liquid. The position of the fragile substance in the liquid is not particularly limited, but is arranged at a position where the lid member does not come into contact with the fragile substance in a state where the lid member is attached to the container to form a closed space (or even if the fragile substance is in contact with the container). Good). Preferably, the frangible is located in the liquid of the container, on the bottom surface of the container, near the bottom surface, or the like.

In the present invention, “water-repellent treatment” refers to processing a solid surface such that a contact angle, which is an angle formed between a liquid surface and a solid surface, is 90 ° or more at a boundary line where the solid surface and the liquid contact. That means. For example, on a water-repellent solid surface treated with Teflon (registered trademark) or the like, the contact angle is close to 180 °, and the droplet becomes almost spherical. The surfaces of the solids do not stick to each other even if a liquid is interposed in the liquid. Examples of the water-repellent processing include a coating method in which a resin such as Teflon (registered trademark), silicon, a fluorine-based water-repellent, wax, alumina soap, pyridinium salt, and seratin is adhered to a solid surface. Further, a method of performing unevenness processing on the solid surface without using a coating agent, such as forming unevenness on the solid surface by laser processing or the like, such as super water repellent processing, may be used.

環状 In the present invention, the term “annular member” refers to a member having a space inside a ring formed by rotating a rectangle around an axis parallel to one side thereof. The height of the annular member is not particularly limited so that the liquid and the fragile substance can be accommodated in such a space, but is typically 1 to 20 mm, preferably 3 to 15 mm, and more preferably 5 to 10 mm. . The inner diameter of the annular member is not particularly limited, but is typically 1 to 15 cm, preferably 3 to 15 cm, and more preferably 3 to 10 cm.

In the present invention, the seal member refers to a member interposed between the container and the lid member to enhance the hermeticity of the container, such as a packing ring.
The annular member and / or the sealing member can be made of an elastic material in order to absorb manufacturing tolerances of the container and the lid member and to enhance the hermeticity of the container. Examples of the elastic material include, but are not limited to, natural rubber, elastomer, nitrile rubber, silicon rubber, and the like.

[First Embodiment]
One aspect of the present invention is a device used detachably mounted on a container for accommodating an implant, a lid member for sealing the container, and a device interposed between the lid member and the container. An annular member for forming a space, wherein the second space is formed between the lid member and the inner surface of the container.

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 the device 1 according to the first embodiment of the present invention, and FIG. 2 is a sectional view of the device 1 shown in FIG. In each of the drawings in the present application, the size of each member is appropriately emphasized for ease of description, and the illustrated members do not indicate the actual size.

As shown in FIG. 1, the device 1 according to the first embodiment of the present invention includes a lid member 3 for sealing a container 2, an annular member 4, a protective member 5, and a seal member 6. The container 2 is a commercially available petri dish having an edge 21 surrounding the opening, and constitutes a storage space in which the liquid L, the fragile material S, and the like can be stored. The lid member 3 includes a top part 32, a convex part 31 extending downward from the top part 32, and a cylindrical skirt wall 34 hanging down from the periphery of the top part 32. The convex portion 31 has a bottomed cylindrical shape having a bottom surface 33 and protrudes toward the internal space of the container 2 with the lid member 3 attached to the container 2. The outer diameter of the protrusion 31 is smaller than the inner diameter of the container 2, and a space G (second space) is generated between the protrusion 31 and the inner surface of the container 2 with the lid member 3 attached to the container 2. Is configured. The cylindrical skirt wall 34 has an inner diameter larger than the outer diameter of the container 2, and functions as a guide mechanism for guiding the position of the projection 31 to the center of the container 2 when attaching the lid member 3 to the container 2.

The annular member 4 is a member having a space (first space) inside a ring formed by rotating a rectangle around an axis parallel to one side thereof. The outer diameter of the annular member 4 is substantially the same as or slightly larger than the inner diameter of the container 2, and is configured to be able to be placed on the bottom surface of the container 2 without displacement. Further, the inner diameter of the annular member 4 is smaller than the outer diameter of the convex portion 31, and when the convex portion 31 is placed on the annular member 4, the convex portion 31 does not enter the space inside the

annular member

4, 4 is configured to be placed on the upper surface. The cross section of the annular member 4 is rectangular, and is configured such that there is no space into which bubbles or the like enter when the annular member 4 is immersed in a liquid. The annular member 4 is made of an elastic material, and when sandwiched between the lid member 3 and the container 2, is configured to absorb manufacturing tolerances and the like of the container 2 and / or the lid member 3 to increase the adhesion. Have been.

The protection member 5 is a member that is placed on the annular member 4 to protect the space inside the annular member 4. The diameter of the protection member 5 is larger than the inner diameter of the annular member 4, and when the protection member 5 is placed on the annular member 4, the protection member 5 does not enter the space inside the annular member 4, It is constituted so that it may be put on. The protection member 5 is made of a fluid-impermeable material so that gas and liquid do not permeate. The protection member 5 is configured as a flexible transparent film-shaped member, and has a sufficient strength that it is not damaged even if it is rolled, bent, or spread. The seal member 6 is an annular elastic member for increasing the adhesion by being interposed between the end 21 of the container 2 and the top 32 of the lid member 3. The inner diameter of the seal member 6 is larger than the inner diameter of the convex portion 31, and the outer diameter is slightly larger than the inner diameter of the cylindrical skirt wall 34. It is configured to be sandwiched between the portion 32 and the edge 21.

As shown in FIG. 2A, when using the device 1, the annular member 4 is placed on the bottom surface of the container 2 containing the liquid L and the fragile object S, and the liquid L and the fragile object S are transferred to the annular member 4. Into the space inside the space (first space). When the liquid volume of the liquid L is lower than the height of the annular member 4, the liquid volume is adjusted such that the liquid volume is equal to or slightly higher than the height of the annular member 4 by pouring the liquid further. Since the cross section of the annular member 4 is rectangular and there is no space for storing gas, the first space is filled with the liquid L. Next, the liquid surface of the liquid L contained in the first space is covered by covering the protective member 5 so as to cover the opening of the annular member 4. Since the protective member 5 is in the form of a film, the protective member 5 can be covered without undulating while gradually increasing the contact area with the liquid surface. Further, since the protective member 5 is transparent, the presence or absence of air bubbles can be visually confirmed, and the air bubbles can be easily pushed out by pressing the protective member 5 with a finger or the like. Thereby, a complete liquid-tight space surrounded by the bottom surface of the container 2, the annular member 4 and the protection member 5 can be formed.

Next, as shown in FIG. 2B, the cover member 3 is attached to the container 2, and the protection member 5 is sandwiched (fixed) between the convex portion 31 and the annular member 4, so that the protection member 5 is shifted from the liquid level. Not to be. At this time, even if air bubbles enter between the convex portion 31 and the protection member 5, the air bubbles do not enter the first space because the protection member 5 is fluid-impermeable. The liquid L extruded by the convex portion 31 is held in the space G between the convex portion 31 and the container 2, and the space between the edge 21 of the container 2 and the top portion 32 of the lid member 3 is Since the hermeticity is enhanced through the seal member 6, even if the liquid L held in the space G moves during the transfer of the device 1, problems such as leakage to the outside of the container 2 hardly occur. The first space is in a completely liquid-tight state and the liquid L is incompressible. Therefore, even if the device 1 vibrates during transfer, the liquid L does not move even if the device 1 vibrates. Does not occur.

When using the fragile material S, the lid member 3 is removed from the container 2 at the transfer destination. However, since the device 1 does not have a structure in which the convex portion 31 enters the liquid-tight space (first space), an operator cannot use the device. It is difficult for the lid member to be removed by a strong force. Further, since the liquid surface of the liquid-tight space is protected by the protection member 5, the liquid surface is less likely to undulate when the cover member 3 is removed from the container 2, and the fragile object S is unlikely to be damaged. Since the film-like protective member 5 is stuck to the liquid L by surface tension, sticking to the protrusion 31 is unlikely to occur. The protective member 5 can gradually reduce the contact area with the liquid surface by gripping and winding the end portion with tweezers or the like, and can minimize the waving of the liquid surface. Then, the fragile object S stored in the first space is taken out and used.

As described above, according to the device 1 according to the first embodiment of the present invention, a liquid-tight state can be efficiently achieved with a simple mechanism and a simple operation, so that there are great advantages in terms of workability and manufacturing cost. is there. And since the inside of a container can be made into a completely liquid-tight state, air bubbles (gas) do not enter in a container and bubbles do not move in a container due to shaking of the container during transfer, and the fragile object is not damaged. In particular, when the fragile material is a laminate of sheet-shaped cell cultures, bubbles do not move, and the laminate does not shift or lose. Therefore, it can be stored for a long time while maintaining the shape of the fragile object in the liquid to prevent deformation.

Further, according to the device 1 according to the first embodiment of the present invention, since the liquid-tight state can be easily released, for example, the vibration of the container or the flow of the liquid due to the forcible removal of the lid member by the operator. Can be suppressed, and fragile objects are not damaged. In addition, since work can be performed without contaminating the surroundings, cleanliness is strictly controlled, such as in a bio-clean room used for producing sheet-shaped cell cultures and in intensive care rooms where sheet-shaped cell cultures are used. Suitable for use in controlled locations.

As described above, the device 1 according to the present invention has been described based on the first embodiment. However, the present invention is not limited to this, and can have various modifications. For example, the protective member 5 and / or The lid member 3 may be subjected to a water-repellent treatment. That is, the liquid L enters between the protective member 5 and the lid member 3 by applying a water-repellent treatment to a portion where the protective member 5 and the lid member 3 are in contact, for example, the bottom surface of the convex portion 31 of the lid member 3. Even if they do not stick. Further, the protection member 5 can be omitted. That is, since the device 1 of the present invention does not have a structure in which the convex portion 31 of the lid member 3 enters the space (liquid-tight space) inside the annular member 4, it is difficult to apply an excessive force when removing the lid member 3. Even if the protective member 5 is omitted, problems such as waving hardly occur. Then, by applying a water-repellent treatment to the bottom surface of the convex portion 31, it is possible to prevent the liquid surface from being pulled by the convex portion 31 and to suppress the waving.

Further, the seal member 6 can also have various modifications. For example, the upper surface of the seal member 6 may be fixed to the top portion 32 of the lid member 3 and a circular groove may be provided on the lower surface side. Thereby, when the lid member 3 is attached to the container 2, the edge 21 of the container 2 enters the circular groove, so that the airtightness of the second space is further improved and the lid member 3 is separated from the container 2. It is hard to come off. At the same time, since the sealing member 6 pulls the lid member 3 downward, the force for pressing the convex portion 31 of the lid member 3 against the annular member 4 is increased, and the liquid tightness of the first space is further improved. Further, the seal member 6 can be freely provided by, for example, being provided between the cylindrical skirt wall 34 of the lid member 3 and the outer peripheral surface of the container 2 or between the convex portion 31 of the lid member 3 and the inner peripheral surface of the container 2. Additions and changes can be made.
Furthermore, the annular member 4 can also have various modifications. For example, when the container is a commercially available petri dish, the inner peripheral surface of the petri dish is slightly inclined. Therefore, by forming at least the outer peripheral surface of the annular member 4 into a tapered shape, the annular member 4 can be configured to be in close contact with the inner peripheral surface of the petri dish. In this case, a mark such as a marker or a small protrusion (potch) may be provided on the upper surface of the annular member 4 so that the upper and lower sides of the annular member 4 can be distinguished. In the case of a protrusion, the top and bottom can be distinguished simply by touching with a hand. However, if the protection member 5 is disposed on the protrusion, the protection member 5 may float up. By providing the notch, it is possible to prevent this floating.
In the present invention, each configuration can be replaced with any configuration that can perform the same function, or any configuration can be added.

The use of the device of the present invention can be performed sequentially, for example, by the following steps.
(1) Prepare a container containing a fragile substance and a liquid.
(2) Place an annular member on the bottom of the container.
(3) Place the protection member on the annular member.
(4) The lid member is attached to the container to form a liquid-tight state.
(5) Transfer the device.
(6) Remove the lid member.
(7) Remove the protection member to release the liquid tight state.
(8) Take out the fragile material and use it.

Hereinafter, an embodiment of a holding device suitable for pressing and holding the device 1 (the container 2 and the lid member 3) of the present invention will be described in detail with reference to the drawings.
FIG. 3 is a perspective view of the holding device 7 according to the second embodiment of the present invention, FIG. 4 is a perspective view of a holding device 7A according to the third embodiment of the present invention, and FIG. 5 is a fourth embodiment of the present invention. It is a perspective view of the holding tool 7B which concerns on a form. In each of the drawings in the present application, the size of each member is appropriately emphasized for ease of description, and the illustrated members do not indicate the actual size. Also,

[Second embodiment]
As shown in FIG. 3, the holding device 7 according to the present embodiment includes a base member 8 and a cover member 9. The holding device 7 can press and hold the device 1 including the container 2 (not shown) and the lid member 3 (not shown) for sealing the container 2 from above and below.

Both the base member 8 and the cover member 9 have circular shapes, and can appropriately press the device 1 from above and below. The base member 8 includes an opening portion 81, a bottom portion 82, a screw portion (engaging portion) 83, a mounting portion 84, and a ridge portion 85. In the present embodiment, the engagement is a screw engagement. The mounting portion 84 has a mounting surface on which the container 2 can be mounted on an upper surface, and an opening 81 is provided at a central portion. The mounting portion 84 has an annular ridge 85 (elongated ridge) surrounding the lower end of the container 2 when the container 2 is mounted on the mounting surface, and defines an installation position of the container 2 with respect to the base member 8. be able to. The mounting portion 84 has a disk shape protruding upward from the bottom portion 82, and a screw portion 83 is provided on a side peripheral surface thereof. The bottom portion 82 has an outer diameter larger than the outer diameter of the mounting portion 84, and serves as a bottom portion when the base member 8 is installed on a table or the like.

The cover member 9 includes an opening 91, a top part 92, a screwing part (engaging part) 93, a cylindrical skirt wall 94, and a grip part 95. The top portion 92 has a cover surface that can cover the upper surface of the lid member 3, and has an opening 91 in the center. The top part 92 includes a tubular skirt wall 94 hanging down from the periphery of the top part 92, and can be sandwiched between the top part 92, the tubular skirt wall 94 and the mounting part 84 so as to surround the device 1. A screw portion 93 (not shown) is provided on the inner peripheral surface of the lower end portion of the cylindrical skirt wall 94, and the screw portion 93 is screwed with the screw portion 83 of the base member 8 to cover the cover. The member 9 can be screwed to the base member 8. Optionally, the inner diameter of at least a portion of the cylindrical skirt wall 94 (for example, the inner peripheral surface at the upper end of the cylindrical skirt wall 94) is matched with the outer diameter of the lid member 3 so that the lid member 3 The installation position can also be defined.

(4) When using the holding tool 7, cover the container 2 in which the storage object S is stored with the lid member 3. At this time, a closed space (liquid-tight space) can be formed by covering the lid member 3 with the container 2 filled with the liquid L arbitrarily. Further, optionally, a sealing member 6 is interposed between the container 2 and the lid member 3 to improve the sealing property, or an annular member is interposed between the container 2 and the lid member 3 as described above. Thus, a liquid-tight space can be formed. Next, the container 2 is placed on the base member 8. At this time, the container 2 can be positioned at the center of the placing portion 84 using the ridge 85. Further, the screw portion 83 of the base member 8 is provided below the mounting portion 84, that is, since the screw portion 83 does not surround the side surface of the container 2, the container 2 is placed on the mounting portion 84. When the container 2 is installed / removed, vibration can be suppressed by sliding the container 2 in the lateral direction. Next, the cover member 9 is attached to the base member 8. At this time, the lid member 3 can be positioned at the center of the cover member 9 (and the base member 8) using the cylindrical skirt wall 94.

As described above, the holding tool 7 can define the relative position between the container 2 and the lid member 3 by using the ridge 85 of the base member 8 and the cylindrical skirt wall 94 of the cover member 9. . When holding the device 1, the screw portion 83 of the base member 8 and the screw portion 93 of the cover member 9 are screwed. At this time, the container 2 and the lid member 3 are vertically connected by gripping a grip portion 95 provided as a protruding portion (may be a groove portion) on the outer peripheral surface of the cylindrical skirt wall 94 and performing screwing work. Can be tightened with strong force. This is suitable for use, for example, when the container 2 is a container having no screwing portion such as a general-purpose petri dish, and has high versatility. In addition, the cover member 3 can be configured not to rotate (screw) with respect to the container 2, and the rotational force of the cover member 3 does not reach the protection member 5 interposed between the container 2 and the cover member 3. It is particularly advantageous. In particular, when the elastic sealing member 6 and the annular member 4 are interposed, these members are accurately compressed and sealed between the container 2 and the lid member 3 whose relative positions are accurately determined. This is advantageous because the property is enhanced. The cover member 9 may be screwed by the height of the screwing portion 83 of the base member 8, so that the clamping operation is easy and the clamping pressure can be easily adjusted. Problems can be prevented.

Since both the base member 8 and the cover member 9 have the opening 81 and the opening 91 at the center, the position where the device 1 is fixed and the liquid L and the stored material S stored in the device 1 are controlled. The state can always be observed. This is particularly advantageous when the container 2 and the lid member 3 are made of a light-transmitting material. Then, even if the graft is a low-visibility object such as a sheet-shaped cell culture, light is emitted from one opening and the sheet-shaped cell culture can be visually recognized from the other opening, which is advantageous. . In the case where the liquid-tight space is formed as described above, the lid member 3 may be pressed against the container 2 while confirming that air bubbles or foreign matter is not mixed into the container 2 through the opening 91. it can. Next, the device 1 is transferred. Since the device 1 is firmly held by the holding device 7, it is possible to prevent a problem that the cover member 3 is detached from the container 2 due to shaking during the transfer and the like. Since it is surrounded by the tool 7, it is protected from external impact. Finally, the holding tool 7 is removed from the device 1 at the transfer destination and used.

Although the present invention has been described with reference to the illustrated embodiment, the present invention is not limited thereto. For example, the threaded portion 83 and the threaded portion 93 are formed by slide engagement (for example, see JP-A-2006-315575), flange fitting (for example, see JP-A-2006-001582), and ridge engagement (for example, JP-A-2006-001582). 2016-203985) or other engaging means having a function of pressing the device 1 from above and below. The annular ridge 85 can be replaced with another means such as a plurality of annularly arranged projections that can position the container 2 on the mounting surface. As the container S, for example, a fragile substance (for example, a laminate of sheet-shaped cell cultures) can be stored. In this case, the container 2 is liquid-tightly transferred while maintaining the shape of the fragile substance. You can also.

[Third embodiment]
Next, a third embodiment of the present invention will be described. Hereinafter, differences from the second embodiment will be described in detail, and description of similar items will be omitted. In each of the drawings in the present application, the size of each member is appropriately emphasized for ease of description, and the illustrated members do not indicate the actual size.

よう As shown in FIG. 4, the holding device 7A according to the present embodiment includes a base member 8A and a cover member 9A. In the present embodiment, the base member 8A further includes a protruding wall 86, a groove 87, and a notch 88. The projecting wall 86 is a plurality of walls provided outside the projecting portion 85 and projecting beyond the height of the container 2 from the mounting portion 84. A groove 87 is provided on the lower surface of the bottom portion 82 of the base member 8A, and when the base member 8A is installed on a table or the like, it is possible to prevent the base member 8A from sticking to the work surface.

When using the holding tool 7A, the container 2 (not shown) is positioned at the center of the base member 8 using the ridges 85. Next, when covering the container 2 with the cover member 3 (not shown), the cover member 3 can be positioned at the center of the base member 8 by aligning the cover member 3 along the projecting wall 86. That is, the worker can define the relative position between the container 2 and the lid member 3 by using the ridge 85 and the projection wall 86 of the base member 8A. This is particularly advantageous when the annular sealing member 6 or the annular member 4 is interposed between the container 2 and the lid member 3 because these members can be pressed accurately.

Next, the device 1 (not shown) is sandwiched. At this time, the device 1 is provided on the side of the bottom portion 82 of the base member 8A and the grip portion 95 configured as a protruding portion radially protruding from the cover member 9A. By gripping the notched portion 88, the screwing operation can be easily performed. When using the device 1, the screwing of the cover member 9 </ b> A is released and the device 1 is removed from the base member 8 </ b> A. At this time, since the device 1 is exposed from the gaps of the plurality of projecting walls 86, the device 1 is exposed. Easy to grab one. Further, since the plurality of projecting walls 86 are arranged so as to surround the container 2, the lid member 3 can be accurately positioned with respect to the container 2.

[Fourth embodiment]
Next, a fourth embodiment of the present invention will be described. Hereinafter, differences from the second embodiment and the third embodiment will be described in detail, and description of the same items will be omitted. In each of the drawings in the present application, the size of each member is appropriately emphasized for ease of description, and the illustrated members do not indicate the actual size.

挟 As shown in FIG. 5, the holding device 7B according to the present embodiment includes a cover member (not shown) and a base member 8B. The base member 8B includes a mounting portion 84B, a ridge portion 85B, and a protruding wall 86B (the description of other components is omitted). The ridge 85B is arranged in a semi-annular shape on the placing portion 84B. The protruding wall 86B is semicircularly arranged outside the protruding ridge portion 85B, and a cutout (gap) C is provided in a part thereof (two or more protruding walls 86B are semicircularly provided with a gap). May be arranged). That is, in the present embodiment, one half of the mounting surface of the mounting portion 84B (the opposite half of the ridge portion 85B and the protruding wall 86B) is configured as a flat surface.

When using the holding device 7B, the container 2 (not shown) is slid in the lateral direction (the direction of the protruding portion 85B and the protruding wall 86B) from the side opposite to the protruding portion 85B and the protruding wall 86B. And install it. At this time, the container 2 is slid on the flat surface of the mounting portion 84B, so that careless vibration of the container 2 can be suppressed. Next, the lid member 3 (not shown) is attached to the container 2. At this time, the lid member 3 is pressed against the projecting wall 86 </ b> B of the base member 8 </ b> B so that the lid member 3 is accurately positioned on the container 2. Can be positioned. When using the device 1 (not shown), the device 1 is removed from the base member 8A. At this time, the side of the device 1 (container 2 and / or lid member 3) exposed from the cutout C is removed. By pushing in, the device 1 can be removed by sliding on a flat surface. A projection 89 serving as a mark may be provided on the mounting portion 84B on the side facing the notch C so that the sliding direction when the device 1 is slid on the base member 8A is easy to understand. Thus, the device 1 can be accurately positioned on the mounting portion 84B only by sliding the device 1 between the protrusion 89 and the notch C. The device 1 is displaced by arranging the protrusion 89 on the extension of the protrusion 85B so as to face the notch C so that the side surface of the device 1 is locked by the protrusion 89. It can also be excluded.

As described above, according to the clamps according to the first to third embodiments of the present invention, the container and the lid can be efficiently brought into close contact with each other with a simple mechanism and a simple operation. There are great benefits. In addition, since the contents in the container can be visually recognized from the outside, the state of the contents, mixing of foreign matter, and the like can be easily checked. Further, since the relative positional relationship between the container and the lid member can be accurately adjusted, the hermeticity is improved. In particular, when an elastic sealing member 6 or an annular member 4 is interposed between the container and the lid member, these members can be pressed accurately, so that the airtightness is further improved.

According to the holding device of the present invention, since the container and the lid member can be securely brought into close contact with each other and fixed, problems such as the lid member coming off the container during transfer can be prevented. In particular, when the container is a laminate of sheet-shaped cell cultures, the container can be reliably transported in a liquid-tight state, air bubbles enter the container and move, and the laminate is displaced or defective. And can be stored for a long period of time while maintaining its shape to prevent deformation.

The present invention is not limited to the above embodiments, and those skilled in the art can appropriately combine the configurations and shapes of the holding devices according to the above-described first to third embodiments, and have different configurations and shapes. Can be realized. For example, the cover member 9 or the cover member 9A may be combined with the base member 8, the base member 8A or the base member 8B, or the protrusion 85, the protrusion 85B, the protrusion wall 86 or the protrusion wall 86B may be used. It may be combined with the base member 8, the base member 8A or the base member 8B.
In the present invention, each configuration can be replaced with any configuration that can perform the same function, or any configuration can be added.

DESCRIPTION OF SYMBOLS 1 Device 2 Container 21 Edge 3 Cover member 31 Convex part 32 Top part 33 Bottom 34 Cylindrical skirt wall 4 Annular member 5 Protective member 6 Seal member

7, 7A,

7B Clamping tool

8, 8A, 8B Base member 81 Opening 82 Bottom 83 Engaging portion 84 Mounting

portion

85,

85B Ridge

86, 86B Projecting wall 87 Groove 88 Notch 89 Projection

9, 9A Cover member 91 Opening portion 92 Top portion 93 Engaging portion 94 Cylindrical skirt wall 95 Grip portion

Claims

A device used by being detachably attached to a container for accommodating a graft, a lid member for sealing the container, and a first space interposed between the lid member and the container to form a first space. The device, comprising an annular member, wherein a second space is formed between the lid member and the inner surface of the container.
The device according to claim 1, further comprising a sealing member interposed between the container and the lid member.
The device according to claim 1 or 2, further comprising a protection member interposed between the annular member and the lid member to protect the first space.
The device according to claim 3, wherein the protection member has flexibility.
The device according to claim 3 or 4, wherein the protective member and / or the lid member are subjected to a water-repellent treatment.
The device according to any one of claims 1 to 5, wherein the graft is a sheet-shaped cell culture.
7. The device according to claim 6, wherein the sheet-shaped cell culture is a laminate.
(8) A holding device including a cover member and a base member configured to hold the device according to any one of claims 1 to 7 from above and below.