JP2003070460A - Backing material for membrane tissue and method for storing/transporting the membrane tissue using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a backing material hard to debond a flat membrane tissue made of a biologically derived material and enabling the membrane tissue to be held advantageously, and to provide a method for easily and inexpensively storing/transporting, packaging and handling, e.g. drawing out, a membrane tissue thus reinforced with such a backing material while preventing the membrane tissue from breakage or the like without impairing its biological properties with its flat shape retained. SOLUTION: In order to storing/transporting this flat membrane tissue made of a biologically derived material, the membrane tissue is lined with the backing material the peripheral rim of which is provided with a means for seizing the peripheral rim.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a backing base material for a membrane tissue and a method for preserving / transporting a membrane tissue using the same, and particularly to a backing substrate capable of advantageously holding a flat membrane-like membrane tissue made of a bio-derived material. Material and the membrane tissue reinforced by such a backing base material, in the state where the flat membrane-like shape is retained, can be preserved or stored for transportation without impairing the biological characteristics of the membrane tissue. To the transportation method.

[0002]

BACKGROUND ART In general, it is applied to defect wound surfaces such as burns, wounds, pressure ulcers, abrasions, skin ulcers or birthmarks, bruise, tattoos, scar removal, laser ablation, etc. to promote healing of the defect wound surfaces. Membrane tissues used for investigating skin irritation, toxicity of chemicals, etc., especially as membrane tissues composed of biological materials, include, for example, skin tissues, mucosal epithelial tissues, corneal epithelial tissues, and cultured skin. , Cultured dermis, cultured epidermal cell sheet, cultured epithelial tissue, cultured corneal tissue and the like, all of which are known to have a very thin film thickness.

More specifically, among those membrane tissues derived from a living body, particularly in the case of a cultured cell sheet-like tissue, the single cell membrane of the cultured epidermal sheet: about 0.01 mm, the single cell membrane of the corneal epithelium: about 0. Even if the culture epidermal sheet has a very small film thickness of 01 mm and is composed of multiple cell layers, the 3-cell layer culture epidermal sheet: about 0.05 to 0.06 mm, the 5 cell layer culture epidermal sheet: about 0. The thickness is 08 mm, and even in the skin tissue (a combination of the epidermis layer and the dermis layer), when the film thickness is thin, it is about 0.2 mm, which is very thin. In addition, since the membrane tissue made of such a bio-derived material has a yuba-like morphology and is not only thin, but also has poor mechanical strength and is very fragile (especially, an epidermal cell sheet made of only cells etc.) In the cell sheet of
It is very difficult to handle), and when they are picked up with tweezers or the like and carried to the defect wound surface, they are easily damaged by their own weight. Therefore, conventionally, in order to protect or retain such a membrane tissue, the membrane tissue is lined with a base material (lining base material) such as gauze or a wound dressing material, and thereby the membrane tissue is reinforced. Although it is easy to handle, even the cultured skin with this backing substrate had a very thin film thickness of about 1.0 mm.

Further, in a facility for culturing and producing such a membrane tissue, a facility for collecting the membrane tissue and performing a treatment such as disinfection, etc., the biological activity and the shape thereof are maintained and the cleanliness is maintained. It was necessary to (sterilely) easily and conveniently package (packaging), store, and transport to a medical institution, and even if the membranous tissue is used in a medical institution, etc. It is necessary to handle tissues cleanly (aseptically) and easily.

By the way, various problems are inherent in fixing and fixing a membrane structure having a thickness of several tens of μm to several hundreds of μm with the above-mentioned backing base material such as gauze or a wound dressing material. is there. For example, as a method for fixing the membrane tissue to the backing base material, a method using a medical stapler, suture, needle or the like is generally considered, and the backing base material is fixed to the membrane tissue according to such a fixing method. Then, since the film structure is likely to be broken from the site of the hole that is necessarily formed in the film structure, it was very difficult to fix the backing substrate to the film structure.
In addition, a method of fixing the membrane tissue to the lining substrate using a medical adhesive (fibrin etc.) has also been proposed, but at the adhesion site, the biological activity of the membrane tissue is lost and surgery is performed. In addition to being difficult to remove before use, there was a problem that it was troublesome and the cost was also very high. There was no choice but to fix the backing substrate with such moisture.

However, in such a method of fixing the backing base material to the membrane structure with water, the membrane structure often spreads surely to the peripheral portion of the backing base material and does not adhere to the backing base material. It was not clear, and it was difficult to use it as a product.
(Because it is necessary to clarify the size when commercializing barley, medical equipment, pharmaceuticals, etc.)

Under such circumstances, under the present circumstances, a flat membrane-like membrane tissue made of a biological material is backed by a base material such as gauze or a wound dressing as described above, so that its biological activity is not impaired. In a commercially available container such as a round petri dish or square petri dish containing a liquid medium, a buffer solution, physiological saline, etc., the lining substrate is housed with the lower side facing down, and a predetermined packaging is performed. It is stored or transported to a medical institution, etc., but the membrane tissue that is only retained on the backing substrate with moisture is liquid medium in the container even if it is wrapped with gauze etc. When exposed to moisture such as, it becomes separated from the backing base material and floats, which causes problems such as the film structure breaking or twisting and the original shape cannot be retained. It is being done.

[0008] Therefore, a storage / transport method that can maintain the original shape and maintain the biological activity without breaking or twisting the membrane tissue before the membrane tissue is used. Development is coveted.

As a container / method for storing or transporting such a membrane tissue, Japanese Patent No. 2904807 discloses that at least a part of constituent materials is gas permeable and bacteria impermeable material. The inner bag and inner bag, and the outer bag made of a metal foil material coated or laminated with a heat-adhesive resin are proposed to have a triple structure, which effectively prevents the problem of microbial contamination. Although it could be resolved, when such a package is used, the artificial skin is still likely to be deformed such as broken or twisted in the inner bag, and the shape of the artificial skin is maintained. The problem that is difficult to solve has not been solved yet. Therefore, the artificial skin is damaged, the biological activity is lowered, and the removal operation thereof cannot be performed with tweezers, which is complicated, and damage is likely to occur during the operation. I have it.

Further, in Japanese Patent Laid-Open No. 8-23968,
Preserving the cultured skin by placing the cultured skin in a gelatin solution that is heated to a temperature, and then lowering the temperature to gel the gel to fix the placed cultured skin. , A transportation method has been proposed, but in the case of a very thin membrane tissue as described above, when simply taking it out from the gelatin gel,
The damage was always caused. In addition, clinically, when applied to humans, there is a risk to the human body of gelatin shock due to residual gelatin.Therefore, after taking it out from the gelatin gel, a very complicated washing operation requiring equipment and personnel is repeated. It is very difficult to handle in a medical field such as an operating room,
It was a method that could not completely eliminate the danger to the human body.

Further, in Japanese Patent No. 2733800, a liquid-tight and airtight package which is generally used for packaging medicines, medical devices, consumer products, etc., more specifically, a flat sheet (cells A skin wound dressing consisting of
A packaging body has been proposed in which a second packaging material is supported on the surface of a bandage support of a liquid-impermeable closed body, which also causes breakage of a cell sheet. The problems such as twisting could not be solved, and the cell sheet clipped to the adhesive substrate sheet [gauze infiltrated with petroleum jelly: petroleum gauze (vaseling gauze)] is still in culture medium during transportation. By the shearing force that oscillates, it falls off from the petroleum gauze, the front and back of the cell sheet become unknown, and it is unknown on which side the layer with germination power to be brought into contact with the wound surface exists.
In addition, there is a possibility that the tweezers cannot be handled and may be damaged.

More specifically, such a patent No. 27338.
In the specification of No. 00, it is believed that the cell sheet and the like can be easily taken out by adhering the cell sheet with an adhesive substrate sheet, that is, petroleum-based gauze (Vaseline gauze) and transporting the cell sheet. In addition, petroleum-based gauze is highly water-repellent and cannot adhere well to cells. Therefore, "the oscillating shearing force of the culture medium during transportation causes the epithelium to fall off the petrolatum-lined lining." The phenomenon was not improved, and in the example, by clipping the cell sheet with petroleum-based gauze,
This prevents the cell sheet from peeling off from the petroleum-based gauze described above. In addition, even if peeling does not occur, during transportation, in the solution such as the medium, the central part of the cell sheet floats and cracks, or the cell sheet consisting of cells is very fragile as described above. Therefore, the local mechanical stress of clip fastening is exerted, which causes damage at the clip holding portion,
With the potential for at least partial loss of biological activity, such clip-fastening requires a very fragile cell sheet to be fastened to the adhesive matrix sheet under sterile conditions, which is very cumbersome. It will be a lot of work. Furthermore, the petroleum component of petroleum-based gauze may remain on the cell sheet, which may adversely affect the normal physiological activity and biological activity of the cell sheet.

[0013]

[Means for Solving the Problems] The present invention has been made in view of such circumstances, and the problem to be solved by the present invention is that it is difficult to detach a flat film-shaped membrane tissue made of a biological material, With respect to the backing base material capable of advantageously holding the membrane structure and the membrane structure reinforced by the backing base material, the flat membrane-like shape of the backing base material is maintained and the biological properties are not impaired and the damage is caused. It is an object of the present invention to provide a method for preserving / transporting a membrane tissue, which is capable of suppressing the above-mentioned problems, and is easy to handle, store and transport such as packaging and taking out the membrane tissue, and is inexpensive.

[0014]

As a result of intensive studies to solve such a problem, the inventors of the present invention have found that, as a flat membrane-like membrane tissue made of a bio-derived material, for example, a culture epidermal cell sheet backing substrate. When fixing or backing, it is possible to utilize the contracting action of the cultured epidermal cell sheet that occurs when peeling the cultured epidermal cell sheet from the bottom of the culture flask, and at least the peripheral portion of the backing substrate. By providing the locking means to at least the peripheral portion of the membrane tissue is locked by the locking means, the backing substrate can be fixedly attached to one surface side of the membrane tissue, It has been found that the membrane structure can be reinforced or retained very advantageously.

Therefore, the present invention has been completed based on such findings, and the gist of the present invention is to dispose on one surface side of a flat membrane-like membrane tissue made of a biological material, A backing base material having a flat plate shape as a whole for holding the film structure, wherein a locking means for hooking and locking at least the peripheral edge part of the film structure is at least the peripheral edge of the base material. A backing base material for a membrane structure, which is characterized in that it is provided in the section.

In short, in the backing base material for a membrane tissue according to the present invention, at least the peripheral portion of the membrane tissue is hooked on the locking means since the locking means is provided at least at the peripheral portion. Thus, the backing base material is advantageously retained by the backing base material. Therefore, as compared with the conventional method of fixing the backing substrate with water such as medium and buffer solution, the peeling or detachment of the membrane structure from the backing substrate is extremely effectively prevented. I have earned it.

According to one of the preferred embodiments of the backing base material for a membrane structure according to the present invention, it is desirable that the backing substrate has a plane area smaller than the plane area of the membrane structure. As a result, the membrane tissue can be effectively locked to the locking means of the backing base material at least at the peripheral portion thereof.

According to another preferred embodiment of the present invention, the locking means is provided on at least a part of a peripheral portion of a surface of the base material facing the film structure. Is provided so as to incline outward or to project in a direction perpendicular to the surface of the base material, and the peripheral edge portion of the membrane tissue is locked by the locking means, thereby holding the film tissue. Such a configuration will be preferably adopted.

Furthermore, according to another preferred embodiment of the backing substrate for a membrane tissue according to the present invention, the locking means comprises:
By providing at least a part of the peripheral portion of the base material so as to project outward from the peripheral portion and in a direction parallel to the surface of the base material, the peripheral portion of the membrane tissue is locked by the locking means. , In contact with one surface of the membrane tissue,
A structure adapted to retain the membrane structure can be suitably adopted.

Further, according to another preferred aspect of the present invention, the locking means includes at least a peripheral portion of a surface of the base material opposite to a surface facing the membrane tissue. A part is provided so as to incline inward from the peripheral edge or to project in a direction perpendicular to the substrate surface, and the engaging means is wrapped around the peripheral edge of the membrane tissue. By stopping it, the structure in which the membrane tissue is held in a state of being in contact with one surface of the membrane tissue is suitably adopted.

In addition, according to another preferred embodiment of the present invention, it is desirable that a liquid passage hole that allows a liquid to pass therethrough is provided through the base material. During storage or transportation of the membrane tissue, it becomes easy to constantly supply the preservation / transport solution to the membrane tissue, so that the cells constituting the membrane tissue are killed or pores are formed in the membrane tissue. The occurrence of such problems can be effectively prevented, and the biological properties (biological activity) of the membrane tissue can be advantageously retained.

As the backing substrate for the membrane tissue according to the present invention, a sheet-shaped, knitted cloth-shaped, non-woven-shaped, or sponge-shaped form made of a bio-derived polymer or a synthetic polymer is advantageously adopted. Can be done.

Further, the present invention relates to a method of storing or transporting a membrane tissue using the above-mentioned membrane tissue backing substrate, wherein the peripheral portion of the membrane tissue is engaged with the locking means of the backing substrate. By stopping and arranging the backing base material fixedly on one surface side of the membrane tissue, the membrane tissue is lined, and the lined membrane tissue is placed in a predetermined liquid-tight container. A membrane tissue, which is characterized by being stored or transported in a state in which the membrane tissue is housed and the membrane tissue is sufficiently moistened with a preservation / transportation liquid while maintaining the flat membrane-like shape of the membrane tissue. The storage / transportation method is also the main point. That is, in the method for storing / transporting a membrane structure according to the present invention, since the backing base material for a membrane structure having the above-described structure is used, the membrane structure is peeled or detached from the backing base material. This can be prevented very effectively and, therefore, even in the preservation / transport solutions used to preserve the biological activity of membrane tissue,
It is possible to avoid as much as possible that such a membrane structure floats, breaks, twists, or the like and cannot maintain the flat membrane shape. Therefore, the flat membrane shape of the membrane structure can be advantageously retained.

Further, in the storage / transportation method according to the present invention, the lined membrane structure is placed in a predetermined liquid-tight container.
From its storage with the storage / transport solution, the membrane tissue can be kept moist at all times, favoring biological activity without compromising the biological properties of the membrane tissue.

In addition, unlike the above-mentioned conventional method, since it does not fix the membrane tissue by using a medical adhesive (fibrin etc.), it is excellent in economy.
Further, since it is not intended to fix the membrane tissue using a gelatin solution or the like, there is no attachment of substances unnecessary for the living body to the membrane tissue, and the packaging and removal of the membrane tissue, The handling operation such as application to the affected area can be easily performed.

According to one of the preferred embodiments of the method for preserving / transporting a membrane structure according to the present invention, the membrane structure backed by the backing substrate has a membrane structure above the backing substrate. It is desirable that the membrane be placed in a position such that the biological tissue characteristics of the membrane tissue are not impaired by the sliding contact between the membrane tissue surface and the mounting surface by adopting such a configuration. Can be
The biological properties (biological activity) of the membrane tissue can be retained to a higher degree.

Further, according to one preferred embodiment of the method for storing / transporting a membrane structure according to the present invention, the membrane structure backed by the backing substrate is placed on a hydrophilic or water-absorbing support substrate. It is possible to preferably employ a configuration in which the predetermined liquid-tight container is housed in the above liquid container. By adopting such a constitution, it becomes possible to make the supporting substrate absorb water or contain water. Therefore, the storage / transport solution can be supplied from the supporting base material to the membrane tissue only by placing the membrane structure on the supporting base material without immersing the membrane tissue in the storage / transporting solution. Therefore, save /
The rocking of the transport liquid causes the lined membrane tissue to collide with and damage the inner wall surface of the liquid-tight container, or the storage / transport liquid scatters when the membrane tissue is taken out from the liquid-tight container. The occurrence of such a problem can be advantageously prevented.

According to another preferred embodiment of the method for storing / transporting a membrane tissue according to the present invention, a holding member is further housed in the liquid-tight container, and the holding member holds A configuration in which the membrane tissue lined by the backing substrate is fixedly held in the liquid-tight container by pressing the membrane tissue lined by the backing substrate is preferably adopted. Can be done. By adopting such a configuration, the membrane tissue contained in the liquid-tight container can be fixedly held in the liquid-tight container by an appropriate pressing force of the pressing member. It is possible to more effectively prevent the tissue from being damaged by being broken, twisted or the like, and it is possible to extremely advantageously maintain the flat membrane-like shape of the membrane tissue.

[0029] In addition, according to another preferred embodiment of the method for storing / transporting a membrane tissue according to the present invention, the holding member and the lined membrane tissue arranged on the supporting base material. It is desirable to have a thickness corresponding to the space between the inner surface of the liquid-tight container facing it,
By adopting such a configuration, it is possible to more effectively suppress the movement of the membrane tissue in the liquid-tight container, even if vibration or impact is applied to such a liquid-tight container,
The flat membrane shape of the membrane structure can be advantageously maintained.

According to another preferred embodiment of the method for preserving / transporting a membrane tissue according to the present invention, the contraction action of the membrane tissue causes the peripheral edge of the membrane tissue to be a means for locking the backing substrate. It is desirable that the peripheral edge portion of the membrane tissue be locked to the locking means at the peripheral edge portion of the backing substrate very easily by adopting such a contracting action of the membrane tissue. ing.

Further, according to another preferred embodiment according to the present invention, the supporting substrate is in the form of a sheet, knitted fabric, non-woven fabric, or sponge made of a bio-derived polymer or synthetic polymer. The configuration exhibiting can be suitably adopted.

In addition, according to another preferred embodiment of the present invention, the membrane tissue is a biological membrane tissue, for example, a mammalian biological membrane tissue such as an epithelial tissue, more specifically a mammal. The skin tissue or epidermis, dermis, mucous membrane or intimal membrane tissue can be exemplified. Examples of the membrane tissue include cultured skin, cultured cornea, or cell sheets such as cultured epidermal sheet, cultured mucosa sheet, and cultured corneal epithelial sheet. By targeting a membrane structure such as these, the features of the present invention can be better exhibited.

According to another preferred embodiment of the present invention, the storage / transport liquid is a liquid capable of maintaining the physiological activity of the membrane tissue, for example, a liquid medium,
A buffer solution, physiological saline, etc. will be used. This allows the biological properties of the packaged membrane tissue to be maintained for an even longer period of time.

Furthermore, according to another preferred embodiment of the storage / transportation method according to the present invention, the liquid-tight container comprises:
A blister case, a tight box, or a lid screwing type case can be suitably adopted. This makes it possible to keep the membrane tissue in a moist state and to advantageously avoid the leakage of the storage / transport liquid contained in the container and the risk of microbial contamination.

[0035]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, various embodiments according to the present invention will be described in detail with reference to the drawings in order to more specifically clarify the present invention. In addition, in FIG. 1 and FIGS. 2, 4, 7, 8, 11, and 12, which will be described later, their thicknesses and the like are exaggerated so that the states of the backing base material and the film structure are exaggerated so that they can be easily understood. It should be understood that is also shown in large proportions.

First, in FIGS. 1A to 1C, a plan view, a side view and a longitudinal section of a flat plate-like backing substrate 10, which is one specific example of the backing substrate for a membrane structure according to the present invention, are shown. The illustrations are each shown schematically. There
The backing substrate 10 is used to support or reinforce the membrane structure described later and maintain its flat film-like shape, and has such a degree as to sufficiently support or reinforce the membrane structure. A material made of a material that has strength and has good adhesion to such a membrane tissue and good biological safety against it (does not adversely affect the biological properties of the membrane tissue) is preferable. Can be adopted.

Specifically, for example, collagen, chitin, chitosan, polylactic acid, polyglycolic acid, polylactic acid-polyglycolic acid mixture, etc., alone or in combination with two of them.
Bio-based polymeric materials using mixtures of two or more species, polyethylene, polypropylene, polyethylene terephthalate, polyacrylates, polymethacrylates, polycarbonates, polyesters, polyvinyl alcohol, polyHEMA (poly-2-hydroxyethylmethacrylate), polyacrylamide, poly Exemplifying a substrate processed into a sheet-like, knitted fabric-like, non-woven fabric-like or sponge-like form made of a synthetic polymer material using N-vinylpyrrolidone or the like alone or a mixture of two or more thereof. Can be done. In addition to the above, it is also possible to employ, as the backing substrate 10, a material having a hygroscopic property or a moisture permeable property, such as cellulose, paper, gauze, or dried animal skin.

The backing substrate 10 made of the above-mentioned materials and forms has a smaller plane area than the film structure to be lined, and as shown in FIG. A base material body 12 having a shape, and the base material body 1
Each of the side portions 2 has a mountain-shaped projection 14 as a locking means, which is integrally provided so as to project outward on the same plane as the base body 12.

More specifically, the chevron-shaped projection 14 has a triangular shape, and the base body 12 is so arranged that one apex 16 of the triangle serves as a side protruding end of the backing base 10.
Is projected outward from each side of the
Since a plurality of such mountain-shaped protrusions 14 are continuously provided on each side of the base material body 12, the outer peripheral edge portion of the backing base material 10 has a sawtooth shape including a mountain portion and a valley portion. It has a tooth shape. In addition, in FIG.
The protruding end of the chevron-shaped projection 14 is sharp because it forms one corner of a triangle, but such a tip (1
6) does not need to be sharp at all, and in order to prevent the damage of the membrane tissue more effectively, a shape in which the corner is taken and the protruding end portion is rounded can be suitably adopted.

Further, in the base material body 12 of the backing base material 10, circular liquid passage holes 18 penetrating in the thickness direction [vertical direction in FIGS. 1 (b) and 1 (c)] are formed at predetermined intervals. The holes are formed in a zigzag pattern, and the liquid can be supplied to both sides of the backing substrate 10 through the liquid passage holes 18. In addition, such liquid passage hole 1
No. 8 is not particularly limited as long as it has a size capable of sufficiently supplying the liquid to the membrane tissue to be lined, but uniform liquid passage to the entire membrane tissue and handling In order to ensure good drainage in
In the range where the intensity of 0 is not reduced extremely,
As a whole, a large area is preferable.

Then, with such a backing substrate 10,
Backed by a flat membrane structure made of a biological material,
Although it is to be supported or reinforced, the membrane tissue to be lined is such as burns, wounds, pressure ulcers, abrasions, skin ulcers or birthmarks such as bruise, tattoos, after scar removal and laser It is applied to the surface of defective wounds after ablation, etc., and is used to accelerate the healing of the surface of the defective wounds, or to examine the irritation to the skin, the toxicity of chemicals, etc. A film-like film structure can be exemplified.

Specifically, as such a membrane structure,
For example, biomembrane tissue, specifically, skin tissue or epidermis,
Dermis, mucous membrane or intimal tissue, mucosal epithelial tissue, mammalian biological membrane tissue such as corneal epithelial tissue, and cultured skin obtained by culturing such biological membrane tissue, cultured dermis, cultured epidermis, cultured mucosal tissue, Examples thereof include cultured epithelial tissue and cultured corneal tissue. The term "cultivated skin" as used herein means that epidermal cells [keratinocytes (keratinocytes)] obtained from mammalian epidermis are cultured,
Proliferated epidermal cell sheet, epithelial cell sheet obtained by culturing and proliferating mucosal epithelial cells obtained from mucosal epithelium, bio-derived polymeric material such as collagen or fibrin after culturing and proliferating fibroblasts obtained from dermis , Laminin, gelatin-like cultured dermis-like ones, and cultured skin-like cultivated dermis-like ones with cultured epidermal cells incorporated therein are known as cultured skin. Further, the cultured corneal tissue refers to a corneal epithelial sheet obtained by culturing and proliferating corneal epithelial cells obtained from the cornea.

Since the various membrane tissues as described above are very thin and fragile like Yuba, it is necessary to handle them with extreme caution. In particular, especially in the case of a membrane structure having a thickness of less than about 0.1 mm, it is difficult to handle it freely when it is operated with tweezers, and it is easily broken, twisted, or damaged. Therefore, it is difficult to maintain the original shape (flat membrane shape), and further, there is an inherent possibility of adversely affecting biological activity. Therefore, the membrane structure as described above is required to have a lining.

By the way, in order to line and fix the above-mentioned film structure with the above-mentioned backing base material 10, the following method can be given as a specific example. In addition, below, as the membrane tissue 22, among the above-mentioned membrane tissues, a cultured membrane tissue obtained by culturing a biological membrane tissue will be described as an example. Incidentally, such a culture membrane tissue can be obtained by a conventionally known method, for example, the method of Green and Rheinwald (US Pat.
16036, Japanese Examined Patent Publication No. 57-18863), etc., and as shown in (a) and (b) of FIG.
The application surface (basal cell layer) on the side of adhering to a biological tissue such as a wound surface is obtained in a state where it is relatively firmly attached.

At this point, first, as shown in FIG. 2A, the upper part of the culture flask 20 is opened so that the operation such as taking out the membrane tissue 22 can be easily carried out. In addition, in the culture flask 20, the upper part of the culture flask 20 is opened by releasing the bonding between the film-shaped lid 26 that closes the upper opening 24 of the culture flask 20 and the culture flask body 27. It can be punished. Then, the backing substrate 10 having a plane area smaller than that of the membrane tissue 22 is put into the culture flask 20 through the opening 24 of the culture flask 20, and the upper surface of the membrane tissue 22 (the surface opposite to the application surface) is centered. Placed in the club.

On the other hand, the membrane tissue 22 is replaced with the culture flask 20.
An enzyme (dispase) solution 28 is added to the inside of the culture flask 20 in order to separate it from the bottom surface 21 of the. Then, this dispase solution 28 is added, and after a while, the membrane tissue 22 starts to peel off from around the peripheral edge [see FIG. 2 (a)], and at the same time, the peeled portion shrinks. The membrane tissue 22 is formed in a state in which it is attached to the bottom surface 21 of the culture flask 20.
This is because the adhesion between the bottom surface 21 and the bottom surface 21 is released by the dispase solution 28, so that the pulled state is released.

By utilizing the contraction action of the film structure 22 as described above, the peripheral edge of the film structure 22 is hooked on the chevron-shaped projections 14 of the backing substrate 10 to be locked. The tissue 22 is lined, and thus the backing substrate 1
The membrane tissue 22 is effectively retained at 0 [see FIG. 2 (b) and FIG. 3]. In addition, in order to lock the peripheral edge of the membrane tissue 22 to the backing base material 10 in this manner, in addition to utilizing the contraction action of the membrane tissue 22, a finger or an auxiliary device such as tweezers is used. It is also possible to lock it.

As described above, in the present embodiment, since the film structure 22 is engaged with the mountain-shaped projections 14 of the backing base material 10, the film structure 22 is peeled or detached from the backing base material 10. The separation can be prevented very effectively. Also, because it does not use medical staplers, sutures, clips, etc.,
Occurrence of damage to the membrane tissue 22 can be advantageously prevented, and
The flat membrane shape of the membrane structure 22 can be held very effectively. Further, as shown in FIG. 3, the film structure 22 lined with the backing base material 10 has the outer shape of the backing base material 10 from where the film structure 22 surely spreads to the peripheral portion of the backing base material 10. It also has the advantage that it exhibits a flat film-like shape having a size substantially the same as the above, and the size becomes clear.

Thus, the above-mentioned dispase solution 28
At, the membrane tissue 22 from the bottom surface 21 of the culture flask 20,
When completely peeled off, the dispase solution 28 is sucked and removed by an appropriate suction tool 30, as shown in FIG. Then, the membrane tissue 22 lined with the backing substrate 10 is transferred from the culture flask 20 to the opening 2 thereof.
It is taken out through 4.

The film structure 22 thus taken out
Is to be put into a predetermined container and packaged. As a container for accommodating such a membrane tissue 22, it is necessary to keep the membrane tissue 22 in a wet state. It is necessary to select a container (liquid-tight container) that can realize liquid-tightness, and by adopting such a liquid-tight container, the biological activity of the membrane tissue 22 can be maintained at a sufficiently high level. It will be. Also,
More preferably, it is desirable to select a liquid-tight container that achieves airtightness to the extent that microbial contamination such as viruses is not caused.

In this embodiment, the culture flask 2
As shown in FIGS. 4 and 5, the membrane tissue 22 taken out from 0 is liquid-tightly and airtightly sealed by sealing the container body 32 and the lid 34 with the annular sealing member 36. It will be accommodated in the liquid-tight container 38.

Here, the container body 3 of the liquid-tight container 38
2 has a box-like shape including a bottom wall portion having a rectangular corner chamfered, and a rectangular tubular side wall portion integrally standing upright from the bottom wall portion at a predetermined height. The lined membrane tissue 22 and the like can be accommodated through the upper opening 40.

Further, one end side of the container body 32 (see FIG. 4,
An engaging portion 44 for engaging the claw portion 42 provided on the lid 34 is integrally provided on the upper side (left side in 5). Furthermore, a shaft member 46 extending in the width direction (direction perpendicular to the paper surface of FIGS. 4 and 5) is immovably fixed to the upper portion on the other end side (right side in FIGS. 4 and 5) of the container body 32. There is.

The shaft member 46 pivotally supports one end of the lid 34 so that the lid 34 can rotate around the shaft member 46 and cannot be pulled out. As a result, the lid 34 and the container body 32 are separated from each other. They are connected at one end (right side in FIGS. 4 and 5). Thus, the lid 34 is attached to the container body 32.
By rotating around the shaft member 46 so as to close the upper opening 40 of the container, and engaging the claw portion 42 provided on the lid 34 with the engaging portion 44 of the container body 32, The container 38 is designed to be liquid-tightly and airtightly closed and polymerized.

By the way, in the present embodiment, when the above-mentioned membrane tissue 22 is accommodated in the liquid-tight container 38 as described above, the inside of the liquid-tight container 38 is kept so that the membrane tissue 22 can be kept in a wet state. A storage / transport liquid 50, which is composed of a liquid capable of maintaining the physiological activity of the membrane tissue 22, such as a nutrient medium (liquid medium), a buffer solution, or a physiological saline solution, is contained in the appropriate amount, and the storage / transport liquid 50 is stored. In order to prevent the storage / transportation liquid 50 from splashing when the lid is opened, a hydrophilic or water-absorbing supporting substrate 52 capable of absorbing or containing the storage / transport liquid 50 is accommodated, and such hydrophilic or The membrane tissue 22 is arranged and supported on the water-absorbent support base material 52.

Here, as the hydrophilic or water-absorbing support substrate 52, it is necessary to select a material that does not reduce the biological activity of the membrane tissue.
Bio-derived polymers such as agar, agarose, collagen, gelatin, alginate, polyvinyl alcohol, and further, hydroxyethyl acrylate, 2-vinylpyrrolidone, dimethyl acrylamide, methyl acrylate, ethyl acrylate, propyl acrylate, butyl acrylate, etc. Examples of the hydrophilic, water-absorbing synthetic polymer obtained by polymerizing monomers singly or in combination, and gel-like, woven cloth-like and non-woven cloth-like substrates formed by singly or in combination can be exemplified. . In addition to these, sponge-like urethane foam, plant fibers, and other conventionally known hydrophilic or water-absorbing substrates can also be advantageously used. Among the materials, those which are not toxic to the cells forming the membrane tissue 22 and are low in cost can be particularly suitably adopted.

Incidentally, the supporting base material 52 in this embodiment.
5A is prepared by pouring the agar sol 54, which is a fluid liquid, into the container body 32 of the liquid-tight container 38 and solidifying (gelling) it, as shown in FIG. Before gelling the agar sol 54,
A support base produced by placing mold materials 58 in the form of swords, in which columnar columnar projections 56 are projected at predetermined intervals, inside the container body 32 and solidifying them, and then removing the mold materials 58. In the material 52 (agar gel 60),
As shown in FIG. 5B, a through hole 62 that can be filled with the storage / transport liquid 50 and accommodated therein is formed.

Then, the membrane structure 22 to which the backing substrate 10 is attached is placed on the supporting substrate 52 (agar gel 60) thus formed, and the supporting substrate 52 is formed.
The storage / transport liquid 50 is added to the liquid-tight container 38 so as to submerge the
By closing and polymerizing the membrane tissue 22, the membrane tissue 22 is packaged while keeping the membrane tissue 22 in a wet state.

When the lined membrane tissue 22 taken out of the culture flask 20 is placed on the support substrate 52, the biological characteristics (biological activity) of the membrane tissue 22 are set to a sufficiently high level. The membrane tissue 22 is positioned above the lining substrate 10 so that it can be held, in other words, the application surface (basal cell layer) to be adhered to the biological tissue such as the wound surface is the top surface. Is desirable.
When the membrane structure 22 is placed below the backing base material 10, the surface of the membrane structure 22 slides on the supporting base material 52,
This is because damage or the like may occur. Even if the membrane tissue 22 is positioned on the upper side, the backing substrate 10 has the liquid passage holes 18 through which the storage / transport liquid 50 can flow, as described above. As shown in FIG.
0 can always be supplied to the entire membrane tissue 22.

As described above, the membrane tissue 22 contained in the liquid-tight container 38 for storage / transportation is generally stored under refrigeration (2 to 9 ° C.) until it is used. Or used for transportation, and before use, at room temperature (19-25
After being left still at (° C.) or kept warm by a heat retaining device (30 to 37 ° C.), it is taken out from the liquid-tight container 38 and used for an operation or a test. Incidentally, such a backing base material 10
Will usually be removed after being used, for example after being applied to the wound surface.

As described above, in the present embodiment, the peripheral edge of the membrane tissue 22 is locked by the locking means (14) provided on the peripheral edge of the backing substrate 10. It is possible to very effectively prevent the film structure 22 from being peeled off or released from the backing base material 10, and therefore, even if it is contacted with or immersed in the storage / transport liquid 50, It is possible to avoid as much as possible that the membrane tissue 22 floats, breaks, twists, or the like and cannot maintain the flat membrane shape. Therefore, in the method for preserving / transporting the membrane tissue according to the present invention, the flat membrane shape of the membrane tissue can be advantageously maintained, and the damage and the like can be effectively prevented.

In addition, in the predetermined liquid tight container 38, together with the hydrophilic or water absorbing support base material 52, the storage / transport liquid 5
From the place where 0 is stored, to the membrane tissue 22,
The storage / transportation liquid 50 is constantly supplied and can be maintained in a wet state, so that the cells constituting the membrane tissue 22 are killed or the membrane tissue 22 is pierced. Can be effectively prevented, and there is no risk of leakage of the storage / transportation liquid 50, whereby the biological activity of the membrane tissue 22 can be extremely improved without impairing the biological characteristics of the membrane tissue 22. And, it can be held for a longer period of time.

Furthermore, since it is not intended to fix the membrane tissue by using an expensive medical adhesive or gelatin solution, it is excellent in economic efficiency and adheres to the membrane tissue, which is unnecessary for the living body. Moreover, it is possible to easily carry out the handling operation such as packaging of the membrane tissue, taking out, application to the affected area, and the like.

By the way, the present invention should not be construed as being limited to only the above-mentioned embodiment, and various modifications can be made without departing from the spirit of the present invention, and other different modifications can be made. Various examples are shown in FIGS.
2 is shown. 6 to 12, members and parts having the same structures as those in the above-described embodiment are given the same reference numerals as those in the above-described embodiment, and detailed description thereof will be given. Will be omitted.

That is, first, in FIGS. 6A to 6C, the membrane structure 22 having a structure different from that of the above-described embodiment is shown.
Another embodiment of a backing substrate for backing is schematically shown. Here, 64 is an upper projecting type backing base material made of the same material and form as the backing base material 10, and is a flat base material having a smaller plane area than the film structure 22 to be backed. It is composed of a main body 66 and a mountain-shaped projection 68 that is integrally provided on the base material main body 66 and that bends and projects upward and serves as a locking means.

More specifically, the backing substrate 64 is
Similar to the backing base material 10, the outer peripheral edge portion thereof has a sawtooth shape including a peak portion and a valley portion, and a part of the peak portion is 90 degrees or more on the upper surface side of the base material main body 66. By being folded back inward at the angle (α), the mountain-shaped projections 68 are provided on the four sides of the base material body 66 so as to incline inwardly from the outer ends and project upward. Of. It should be noted that, like the backing base material 10, the base body 66 of the backing base material 64 has an oval or penetrating shape in the thickness direction thereof (vertical direction in FIGS. 6B and 6C). The slit-shaped liquid passage holes 70 are formed in a zigzag pattern at predetermined intervals.

Then, in order to line the target membrane structure 22 with the backing base material 64 as described above, FIG.
As shown in (a), first, the upper top plate (not shown) of the culture flask 19 is removed to make it open, and then the membrane tissue 22 is put into the culture flask 19 through the opening 25.
The backing base material 64 having a smaller planar area is formed on the surface opposite to the surface opposite to the membrane tissue 22 so that the surface on the side where the chevron projections 68 project is the upper surface. The chevron-shaped projections 68 are allowed to stand so that they are allowed to stand on the central portion of the upper surface (the surface opposite to the application surface) of the membrane tissue 22.

Then, in order to separate the membrane tissue 22 from the bottom surface 23 of the culture flask 19, an enzyme (dispase) solution 28 is added into the culture flask 19. Then, a short time after adding the dispase solution 28, the membrane tissue 22 begins to peel off from the peripheral portion thereof [FIG.
(See (a)], the contraction action of the membrane tissue 22 is used from the beginning of contraction so that the mountain-shaped projection 68 of the backing base material 64 is wrapped around the periphery of the membrane tissue 22. It can be stopped. As a result, the one side of the backing base material 64 is in contact with the membranous tissue 22, and the membranous tissue 22 is lined, so that the flat film-like shape of the membranous tissue 22 can be retained [Fig. 7 (b)].

Then, the membrane tissue 22 lined as described above is taken out from the culture flask 19 in the same manner as in the above-mentioned embodiment, and the membrane tissue 22 can be kept in a wet state. In addition, it is stored in a container (liquid-tight container 72) capable of realizing liquid-tightness as shown in FIG. 8, packaged, and stored / transported.

Here, the liquid-tight container 72 includes a bottom wall portion having a rectangular corner chamfered, and a rectangular tubular side wall portion integrally provided upright from the bottom wall portion at a predetermined height. A container having a so-called blister case structure composed of a container body 74 and a film-like lid body 76 that closes the upper opening of the container body 74, and the lined membrane tissue 22 through the upper opening of the container body 74. Or storage / transport liquid 50,
A supporting base material 78 described later can be housed and enclosed. Further, on the outer periphery of the upper opening portion of the container body 74, a thin flange portion (flange portion) 80 that projects outward by a predetermined dimension and continuously extends in the circumferential direction is integrally provided. On the upper surface of the portion 80,
The upper opening of the container body 74 is covered and closed by being bonded by a conventionally known method, so that the liquid-tight container 72 is liquid-tightly sealed.

By the way, in the liquid-tight container 72 described above,
As shown in FIGS. 8 and 9, the support substrate 52 described above is used.
A supporting base material 78 having a structure different from that of is accommodated. Specifically, the supporting base material 78 is used as the storage / transport liquid 5.
It is composed of a sponge body 82 capable of absorbing or containing 0, and has a substantially rectangular thick plate shape having a lower height than the liquid-tight container 72. Further, in the central portion on the upper surface side thereof, a substantially rectangular recess 84 for mounting the lined membrane tissue 22 is formed.
And side working recesses 86 for facilitating the removal of the membrane tissue 22 are provided in the central portions of the four side walls of the recess 84.

Then, in the recess 84 of the supporting base material 78 (sponge body 82), the membrane structure 22 having the backing base material 64 attached thereto is positioned above the backing base material 64. Liquid-tight container 72
The storage / transport liquid 50 so that the supporting substrate 78 is dipped therein.
Then, the upper opening of the liquid-tight container 72 is closed with a lid 76.
The membrane tissue 22 is kept in a wet state and is provided for storage / transportation by being sealed at.

As described above, according to the present embodiment as well, since the peripheral edge portion of the membrane tissue 22 is locked by the locking means (68), the membrane tissue 22 can be freely detached and moved. Instead, it can be effectively retained.
Therefore, even in the liquid-tight container 72, the film tissue is separated,
It is possible to very advantageously avoid the problems that the flat membrane shape cannot be maintained due to floating, breaking, twisting, etc., and further, the membrane tissue is damaged. Of.

Further, FIGS. 10A to 10C each schematically show one of the backing base materials having a structure different from that of the above embodiment. There
Reference numeral 90 denotes an upper projecting type backing base material having the same material and form as the backing base material 10, and a base material body 92 having a substantially rectangular flat plate shape having a plane area smaller than that of the film structure 22. , A pin-shaped projection 94 as a locking means fixed to the base material main body 92.

More specifically, 4 of such backing substrate 90
A total of 22 pin-shaped projections 94 are formed on the outer peripheral edge portion including the two corner portions, which are composed of a reverse taper portion that increases in diameter upward and a column portion that extends downward from the reverse taper portion at predetermined intervals. The backing base material 90 is fixedly attached at a predetermined interval so as to project from the upper surface of the backing base material 90 in the same direction and in a direction perpendicular to the base material surface. Further, in the base material body 92 of the backing base material 90, as in the backing base material 10, circular liquid passage holes 96 penetrating in the thickness direction thereof are provided at predetermined intervals, and a total of 13 holes are provided. The holes are staggered, and the liquid can be passed through the liquid passage holes 96.

Then, with such a backing base material 90,
In order to line the desired flat membrane-like membrane tissue 22, first, as shown in FIG.
The upper part of the culture flask is opened, and the culture flask 2 is opened through the opening 24.
0, so that the surface on which the pin-shaped projections 94 project is the upper side, in other words, the pin-shaped projections on the surface opposite to the surface facing the membrane tissue 22. The membrane (22) is allowed to stand on the center of the upper surface (the surface opposite to the application surface) of the membrane tissue (22) so that the membrane (22) is detached from the bottom surface (21) of the culture flask (20). The solution 28 is added to the culture flask 20.

Thereafter, the contraction action of the membrane tissue 22 caused by the addition of the enzyme solution 28 is utilized, and it is picked up by an auxiliary device such as tweezers and fixed to the peripheral portion of the backing base material 90. The pin-shaped projection 94 thus formed is wrapped around the peripheral edge of the membrane tissue 22 so that the membrane tissue 2
By hooking 2 and locking it, the membrane tissue 22 is lined. Then, this allows the flat film-like shape of the membrane tissue 22 to be extremely advantageously held in a state where the backing base material 90 is in contact with the membrane tissue 22 [see FIG. 11 (b)]. Since the tip of the pin-shaped projection 94 has a large diameter and is thick, it is even easier to lock the membrane tissue 22.

Thus, the membrane tissue 22 lined as described above is taken out from the culture flask 20 in the same manner as in the above example, and the container (liquid) as shown in FIG. After being housed in a tight container 38) and subjected to packaging, it is provided for storage / transportation.

Also in the present embodiment, as shown in FIG. 12, a supporting base material 98 made of a non-woven fabric capable of absorbing or containing the storage / transport liquid 50 is placed in the liquid tight container 38. In addition, the supporting substrate 98 includes
Similar to the above example, a substantially rectangular recess for mounting the lined membrane tissue 22 and a side working recess for taking out are provided.
It is provided. Then, the film structure 22 having the backing base material 90 attached to the recess of the support base material 98 is
The membrane structure 22 is placed so as to be located above the backing base material 90, and the supporting base material 98 is immersed therein.
The storage / transport liquid 50 is added to the liquid tight container 38.

By the way, in the present embodiment, as described above,
After the membrane tissue 22 is placed on the support substrate 98, as shown in FIG. 12B, the upper surface of the membrane tissue 22 and the inner surface of the liquid-tight container 38 (lid 34) facing the upper surface of the membrane tissue 22. 39
A pressing member 100 having a substantially thick-walled lattice shape, which has a thickness substantially similar to the thickness d corresponding to the space between and, is housed above the membrane tissue 22. The combination of the base material 90 ″ can be fixedly held in the liquid tight container 38. The liquid-tight container 3
After storing the combination as described above, 8 is liquid-tightly sealed as in the previous embodiment.

Therefore, if such a configuration is adopted, the above-mentioned pressing member 100 presses the membrane tissue 22 with an appropriate pressing force that does not damage it, and thus the membrane tissue 22 is With a proper pressing force of the pressing member 100, the pressing member 100 can be fixedly held in the liquid-tight container 38. Therefore, the membrane tissue 22 may be broken,
Twisting or damage such as breakage can be prevented more effectively, and the flat membrane-like shape of the membrane tissue 22 can be retained very advantageously. Therefore, even if vibration or impact is applied to such a liquid-tight container 38, the flat film-like shape of the film tissue 22 can be advantageously maintained.

Here, as the pressing member,
In addition to the pressing member 100 having a substantially thick lattice shape as in the above example, the shape, form, size, etc., of the pressing member 100 as long as the movement of the membrane tissue 22 in the liquid tight container 38 can be restricted, The shape is not particularly limited, and may be, for example, a sheet shape, a flat plate shape, a corrugated plate shape, a sponge shape, or a shape capable of uniformly pressing the membrane tissue 22, such as the liquid-tight container 3.
On the lower surface of a frame body or a plate-like body having substantially the same size as the inner circumference of 8, the protruding body having a thickness substantially similar to the above-mentioned thickness: d is arranged so that the peripheral edge portion or the central portion of the membrane tissue 22 is Examples thereof include a structure that is pressed with a small area, and a needle-shaped object that is pressed with an end portion or the like of the membrane tissue 22. However, it goes without saying that the size and the like of such a pressing member need to be appropriately set so as not to excessively press the membrane tissue 22.

As the material of the pressing member, any of metal, vegetable fiber, biopolymer, and synthetic polymer material can be selected. Among them, water-repellent synthetic polymer material, metal, etc. As described above, a material having a property of being difficult to adhere to the film structure 22 can be particularly preferably adopted. As a result, the occurrence of problems such as the membrane tissue 22 sticking to the pressing member 100 can be advantageously avoided.

As described above, in the embodiment shown in FIGS. 10 to 12, since the peripheral edge portion of the membrane tissue 22 is locked by the pin-shaped locking means (94),
2 is effectively retained without desorption. Further, in the present embodiment, the membrane tissue 22
Of the liquid tight container 3 from the place where the pressing member 100 for suppressing the movement of the liquid tight container 3 in the liquid tight container 38 is housed.
The membrane tissue 22 accommodated in 8 can be fixedly held in the liquid-tight container by an appropriate pressing force of the pressing member, and the membrane tissue is broken or twisted. This can be prevented even more effectively, and the flat membrane-like shape of the membrane structure can be retained very advantageously.

The representative embodiments of the present invention have been described in detail above, but they are merely examples, and the present invention is not limited to the specific description of such embodiments. It should be understood that it should not be construed as limiting.

For example, in the above embodiment, the backing base materials 10, 64, 90 are formed into a substantially rectangular flat plate shape as a whole, whereby the backing base materials 10, 64, 9 are formed.
The film structure 22 lined with 0 was also formed into a rectangular flat film shape, but the shape of the backing base materials 10, 64, 90 is not limited to the illustrated one, and for example, a rectangular shape. It does not matter even if the shape is a combination of a circular shape and an elliptical shape, and in any case, depending on the shape of the film structure before lining, the shape of the film structure desired at the time of use, etc.,
It can be set appropriately. However, so that the backing base material has a smaller planar area than the membrane tissue so that the peripheral edge of the membrane tissue can be locked, the backing base material can be locked at the portion of the membrane tissue protruding from the backing base material. Needless to say, it is desirable to do.

In the backing base material 10, the mountain-shaped projections 14 as locking means are continuously provided on the entire peripheral edge portion, and in the backing base materials 64 and 90, the entire peripheral edge portion is provided. The mountain-shaped projections 68 or the pin-shaped projections 94 as the locking means are provided at a predetermined interval on the backing base material 10, 6 of the locking means.
As long as it can be surely spread and held on the entire one side surface of the base material 4, 90, at least the base material body 12, 6
It is possible to provide them at a part of the peripheral edge portions of 6, 92, for example, in the case of a rectangular backing base material, at the periphery of four corner portions. Further, the number, shape and size of the locking means provided are not limited to those in the above example, and the shape and size of the membrane tissue 22 to be lined,
The thickness can be appropriately set according to the contraction force of the membrane tissue 22 and the like.

For example, in the backing base material 10 provided with the locking means (14) so as to project outward from the peripheral edge portion of the base material and in the direction parallel to the base material surface, the apex of the mountain portion More preferably, that is, it is more desirable to make the apex angle of the crest part obtuse, thereby advantageously suppressing damage to the peripheral edge of the membrane tissue 22 locked by the crest projection 14. Can be done.

Further, the backing base materials 64, 90 provided with the locking means (68, 94) so as to incline inward from the peripheral edge portion of the base material or to project in a direction perpendicular to the base material surface. Then, as the protruding height of the locking means: h, for example,
When handling a membrane tissue having a thickness of 0.1 mm or less, such as a cultured epidermal cell sheet, usually 0.5 to 5 mm
A protrusion height of about h: h can be suitably adopted. If the protrusion height: h is too high, the locking means (6
8) It becomes difficult to lock the membrane tissue 22 in 8), and it may be difficult to cover the transplanted part with gauze or a hydrophilic or hydrophobic wound dressing material, which is usually performed after transplanting the membrane tissue. And vice versa:
If h is too low, the membrane tissue 22 may not be locked well.

Further, as described above, the backing substrate 6
In Nos. 4 and 90, the locking means (68, 94) is provided inwardly from at least a part of the peripheral edge of the surface opposite to the surface facing (contacting) the membrane tissue. It was provided so as to be inclined to or protruding in a direction perpendicular to the surface of the base material, whereby the peripheral edge portion of the membrane tissue 22 could be easily locked. The locking means (68, 94) does not matter even if it is provided on the surface facing the membrane tissue. In the case of adopting such a configuration, the locking means (68, 94) is It is desirable to be provided so as to be inclined outward from the peripheral portion of the base material or to be projected in a direction perpendicular to the surface of the base material. The part can be easily locked. Also,
In particular, when the projection is made in the vertical direction, the locking means (9
It is more desirable to employ a locking means that becomes thicker toward the tip as in 4).

As described above, even when the locking means is provided on the surface facing the membrane tissue 22, the locking means should be provided on at least a part of the peripheral portion of the backing substrate. It is sufficient that the number, shape, and size of the arrangement can be appropriately set according to the shape, size, and thickness of the membrane tissue 22 to be lined, and further the contraction force of the membrane tissue 22. For example, like a cultured epidermal sheet, thickness: 0.1 mm
In the case of lining the following membrane tissues, the protrusion height of the locking means may be preferably about 0.5 to 3 mm. This is because if the protrusion height is too high, the locking means may excessively enter the transplant bed during application (implantation), which may make it difficult to remove. 22 is too empty, and the membrane tissue 22
May not be effectively reinforced and may be easily damaged. On the contrary, when the protrusion height is too low, the membrane tissue 2
This is because there is a possibility that 2 cannot be locked well. Further, in the case where the locking means is provided on the surface facing the membrane tissue 22 as described above, the plane area of the backing base material is not necessarily smaller than the plane area of the membrane tissue to be lined. It doesn't have to be.

As described above, according to the present invention, with respect to the locking means provided on at least the peripheral portion of the backing base material,
By locking at least the peripheral portion of the film structure, the film structure can be reliably spread and held up to the end of the backing substrate. By the way, depending on the material of the backing base material, there are those in which irregularities are accidentally formed on the surface of the base material and projections are formed.However, such an accidentally formed projection has a backing base material, for example. Even if formed on the peripheral portion of the material, since the film structure cannot be satisfactorily locked to such a protrusion, the projection does not surely spread over the entire surface of the base material and the size is not clear. It cannot be the locking means according to. In addition, here, the protrusions formed accidentally refer to, for example, fluffing in a backing substrate made of a non-woven fabric, a woven fabric, or the like, a synthetic polymer or a biopolymer-derived material, and unevenness of the surface of animal skin. There is.

Further, in the above embodiment, the backing substrate 1
0, 64, 90 (base material bodies 12, 66, 92) were provided with liquid passage holes 18, 70, 96, respectively, which allow liquid to pass therethrough.
If the storage / transportation liquid 50 can be supplied to the membrane tissue 22 lined with 4, 90, those liquid passage holes 18,
70 and 96 are not always required. However, due to the liquid passage holes 18, 70, 96,
Needless to say, the distribution of the storage / transport liquid 50 can be further improved. Further, the shapes, opening densities, sizes, positions, and numbers of the liquid passage holes 18, 70, 96 are not particularly limited, and can be appropriately set as necessary. For example, various shapes such as a circular shape, an elliptical shape, a rectangular shape, a polygonal shape, and a linear shape can be adopted, and the strength of the backing base material 10, 64, 90 is extremely weakened even if the opening density and the like are large. It can be set as appropriate to the extent that it does not occur.

In addition, in the above example, as the membrane tissue to be lined, the membrane tissue 22 which has been cultivated and propagated in the culture flask 20 to have a flat membrane shape (sheet shape) has been illustrated. Is by no means limited to such a membrane tissue 22, but any flat membrane-like membrane tissue made of a bio-derived material can be targeted, and in particular, a culture container such as a culture flask or a petri dish. It is desirable to target a membrane tissue cultured in a sheet form, for example, a cultured epidermal cell sheet, a cultured mucosal cell sheet, or a cultured corneal cell sheet. However, since these membrane tissues exhibit a contracting action when the membrane tissues are detached from the culture container by an enzyme such as dispase or a temperature change, a locking means for the backing substrate 10, 64, 90 ( 1
4, 68, 94) is extremely easy to lock.

Further, even in the case of the liquid-tight container, the material, shape, size and the like of the container are not limited as long as the container can realize the liquid-tightness and can accommodate the membrane tissue 22 and the supporting base material. However, various conventionally known ones can be adopted, and it is by no means construed as being limited to the structure of the liquid-tight containers 38 and 72 in the above example. In addition to the above, various conventionally known liquid-tight containers such as an aluminum pack, a plastic case, a blood bag, a tight box, a lid body screwing type case and the like can be adopted. In addition, among such liquid-tight containers,
The bag-like structure has a liquid-tight container 38 because the stored / transported liquid 50 that is enclosed is likely to be scattered or spilled out when the membrane tissue is taken out at a site of use such as an operating room, a treatment room, or a hospital room. Like 72, a blister case structure or a tight box structure composed of a box-shaped container body and a lid body, and further, the lid body is screwed into the container body to close the opening thereof. A liquid-tight container having a lid screw-engagement case structure can be particularly preferably adopted.

Further, in these liquid-tight containers, the movement of the membrane tissue contained in the container is restricted to one or more places on the upper inner surface, the lower inner surface, and the side wall surface to prevent the occurrence of damage or the like. It does not matter at all if the pressing member (protective member) for prevention is fixedly provided. Specifically, for example, in the above embodiment, the pressing member 100 is the liquid-tight container 38.
It may be provided integrally with the lid 34.

Further, in the above embodiment, the membrane tissue 22 taken out from the culture flask 20 was directly stored in the liquid-tight containers 38 and 72 and packaged. However, before packaging, It does not matter even if the membrane tissue 22 is washed. Barefoot, membrane structure,
Since blood components and the like are attached to cell sheets derived from animals, and serum and the like are attached to cultured skin, it is necessary to perform sufficient washing operation during storage. Is.

In addition, the injection amount of the storage / transportation liquid 50 in the above-mentioned embodiment is not limited to the exemplified one, as long as the membrane tissue can be maintained in a wet normal state, and the membrane tissue and the supporting group are not limited thereto. It can be appropriately set according to the material and the like.

Further, the timing of disposing the backing substrate 10, 64, 90 on the membrane tissue 22 in the culture flask 20 is not particularly limited, and for example, for the sheeted membrane tissue 22, Backing substrate 10, 64, 90
Even if the dispase solution 28 is added after the disposing of the diaper, the backing substrate 1
0, 64, 90 may be arranged on the membrane tissue 22.

Although not listed one by one, the present invention is
Based on the knowledge of those skilled in the art, it can be implemented in various modified, modified, and improved modes, and
It goes without saying that all such embodiments are included in the scope of the present invention without departing from the spirit of the present invention.

[0101]

EXAMPLES Hereinafter, representative examples of the present invention will be shown to clarify the present invention in more detail. However, the present invention is not limited by the description of such examples. Needless to say, it is not something to receive.

Example 1 First, the epidermis of a human being, a mammal, was collected and used for the method of Green and Rheinwald (US Pat. No. 4016036, Japanese Patent Publication No. 57-1886).
3), a feeder layer (supporting cell layer) prepared by stopping the growth ability of fibroblasts with mitomycin C and a cell culture medium are preliminarily sealed with a top plate as shown in FIG. Can be peeled off, TPP
It was housed in a culture flask made by (Techno Plastic Products AG). Then, in the culture flask (20),
The epidermal cells [keratinocytes (keratinocytes)] having a proliferation ability obtained from the epidermis were seeded and cultured in the same manner as in the conventional method, and spread over the entire area of about 150 cm ² on the bottom surface (21) of the culture flask (20). Cultured epidermal cell sheet (2
2) was obtained. The cultured epidermal cell sheet (22)
Was formed from two cell layers.

Next, after opening the upper part of the culture flask (20), a dispase (manufactured by Godo Shusei Co., Ltd.) solution (28) was injected to the bottom (2) of the culture flask (20).
The cultured epidermal cell sheet (22) adhered to 1) was peeled from the culture flask (20). At the time of this peeling, FIG.
A polyethylene terephthalate-lined backing substrate (10) having a shape as shown in (1) is placed on the center of the upper surface of the cultured epidermal cell sheet (22), and the cultured epidermal cell sheet (22) is adjusted according to the contraction of the cultured epidermal cell sheet (22). The peripheral edge part of () was hooked on the chevron projections (14) of the lining substrate (10) and retained, so that the cultured epidermal cell sheet (22) was lined. The backing substrate (1
0) is length: 8 cm x width: 6 cm x thickness: 0.2 mm
It is produced by cutting the peripheral edge of the rectangular flat plate-shaped base material so that the pitch (16) pitch of the chevron projections (14) is 3 mm and the projecting length is 2 mm. 14) were provided in the longitudinal direction with 25 pieces each and in the width direction with 18 pieces each. Further, in the base material main body (12) of the backing base material (10), 20 liquid passage holes (18) with a diameter of 3 mm were formed in a zigzag pattern at intervals of 10 mm. Then, the backing substrate (10) was subjected to a predetermined sterilization treatment before use.

Then, after removing the injected dispase (combined liquor purification) solution (28) from the culture flask (20), the lined culture epidermal cell sheet (22) was
It was taken out from the culture flask (20) with tweezers. Then, the thus-obtained cultured epidermal cell sheet (22) is provided with a blister case (72) having a brim (80) around the opening (length: about 100 mm × width: about 90 mm × depth: Approximately 15 mm), the backing substrate (10) was placed on the hydrophilic hydrophilic sponge (82), which is the supporting substrate (78), so that the application surface of the cultured epidermal cell sheet (22) was the upper surface. ) Was attached and the surface was left still.

Thus, the cultured epidermal cell sheet (2
Since the dispase solution (28) used at the time of peeling from the culture flask (20) was attached to 2), a washing operation for removing this dispase solution (28) was carried out. That is, cultured epidermal cell sheet (22)
The culture epidermal cell sheet (22) was washed by repeating the operation of injecting and discharging the phosphate buffer solution into the blister case (72) accommodating the above.

Then, such a blister case (72)
A serum-free green medium was added to the inside to such an extent that the cultured epidermal cell sheet (22) was immersed, and the aluminum film as the lid body (74) was covered over the entire opening of the blister case (72). Thereafter, the film was thermocompression-bonded to the brim case (80) of the blister case (72) to liquid-tightly close the opening of the blister case (72) for packaging. This is 2 ℃ ~
After being stored for 48 hours in a refrigerating environment within a temperature range of 9 ° C., they were transported in the same refrigerating environment. Then, the temperature was returned to room temperature (about 25 ° C.), the bag was opened, and the backing substrate (10) and the cultured epidermal cell sheet (22) were taken out with tweezers. The flat film shape was maintained.

Example 2 Human skin (skin layer + dermis layer) collected in a spindle shape was immersed in a dispase (purified joint liquor) solution for about 2 hours, and then,
The epidermis layer was peeled off. At this time, the temperature of the dispase solution was controlled in a thermostat of 30 to 36 ° C. Then, the obtained human epidermis is immersed in a trypsin solution (manufactured by Lifetech Oriental Co., Ltd.) controlled at 30 to 37 ° C., then centrifuged, and the collected epidermal cells [keratinized cells (keratinocytes)] are fetal bovine. Suspend in a green medium containing 3% serum, inoculate the suspension into a culture flask (20),
Primary culture was carried out in the same manner as in the past. In addition, before seeding the epidermal cells in the culture flask (20),
A feeder layer obtained by treating mouse-derived 3T3 cells with mitomycin C was previously housed. The primary culture was carried out by removing the epidermal cells with a trypsin solution in a subconfluent state, preparing a suspension again, and subculturing in the same manner as the primary culture to form a sheet. . Then, in the subculture, the epidermal cells are in a confluent state, that is, the bottom surface (2) of the culture flask (20).
1) (about 10 cm × about 8 cm), it was confirmed that the cultured epidermal cells were in a sheet form, and then the cells were further cultured for 3 days. The prepared cultured epidermal cell sheet (22) had one layer in the thin portion and three layers of keratinized cells in the thick portion.

On the other hand, as the backing substrate (10), the length:
9 cm × width: 6 cm × thickness: about 0.2 mm, the peripheral edge portion of a rectangular flat plate-shaped chitin nonwoven fabric (Vesquitin W: made by Unitika) was subjected to the same procedure as in Example 1 above, and the apex () of the chevron protrusion (14) ( 16) By cutting so that the pitch is 5 mm and the protruding length is 3 mm, 16 mountain-shaped projections (14) as locking means are provided in the longitudinal direction and 10 in the width direction. Backing substrate according to Example 2 (1
0) was prepared. It should be noted that the backing substrate (10) according to the second embodiment has a substrate body (12) having a liquid passage hole (18) having a diameter of about 2 mm and penetrating in the thickness direction of the substrate (10).
Twelve staggered holes were formed at intervals of 5 mm. Further, the backing substrate (10) was subjected to a predetermined sterilization treatment before use.

Then, after removing the medium in the culture flask (20) and washing the cultured epidermal cell sheet (22) with Hanks' solution, 2 mL of the dispase solution (28) was added and the temperature was raised to 25 to 37 ° C. Culture flask (20)
Was heated, and when a part of the peripheral edge was peeled off, the backing substrate (10) according to Example 2 described above was placed in the center of the cultured epidermal cell sheet (22). Then, from the bottom surface (21) of the culture flask (20) to the culture epidermal cell sheet (2
In accordance with the peeling-off and contraction of 2), the peripheral edge of the cultured epidermal cell sheet (22) is supplemented with tweezers to become a mountain-shaped projection (14) on the peripheral edge of the backing substrate (10). The cultured epidermal cell sheet (22) was lined by hooking and indwelling.

Next, after opening the upper part of the culture flask (20), the dispase solution (28) was removed, and the operation of injecting and discharging the phosphate buffer was repeated several times to obtain a culture flask (20). Inside, the cultured epidermal cell sheet (22) was washed.

On the other hand, as shown in FIG. 5, the lid (3
4) and the container body (32) are liquid-tightly sealed in a liquid-tight container (38) in a fluid state (agar sol 5).
4) was poured and solidified at room temperature. At this time, the columnar projections (56) (diameter: about 1 mm) of the Kenzan-like material (58) made of acrylic resin were placed so as to be perpendicular to the bottom surface of the liquid-tight container (38), and the agar sol (54). By solidifying and removing this mold material (58) after solidification, the agar gel (60) is provided with a through hole (6
2) was produced at intervals of about 10 mm. Then, the medium (50) was further added so that the agar gel (60) thus formed was sufficiently filled with the medium (50).

Then, the cultured epidermal cell sheet (22) lined with the backing substrate (10) was placed on the supporting substrate (52) made of the agar gel (60) with the basal cell layer facing upward. That is, the cultured epidermal cell sheet (22) was arranged so as to be located above the lining substrate (10). Then, by engaging the claw portion (42) of the lid body (34) of the liquid-tight container (38) with the engaging portion (44) of the container body (32), the upper opening of the container body (32). (40) was closed, polymerized, and packaged.

The above operation was carried out in a room temperature environment of about 19 to 26 ° C., and the packaged cultured epidermal cell sheet (22) was kept at a temperature environment of 2 to 9 ° C. for 24 hours. Was stored in a car-mounted electric refrigerator (about 5 to 8 ° C.) mounted on a sponge-like cushioning material having a thickness of 5 cm and transported to a medical institution for about 2 hours. Then, before being used in a medical institution, after being put in a warmer at about 35 ° C., the claw portion (42) of the lid body (34) and the engagement portion (44) of the container body (32) are engaged. Is released, and the cultured epidermal cell sheet (22) in the liquid-tight container (38) is placed on the backing substrate (10) and then the backing substrate (1).
The size was almost the same as that of 0) and the flat film shape was maintained.

Example 3 A peripheral edge of a polyethylene sheet having a length of 9 cm, a width of 6 cm and a thickness of 0.2 mm was processed in the same manner as in the above Examples 1 and 2 so that the apex (16) pitch of the ridges was 5 mm. , Sawtooth-shaped (mountain-valley-mountain-valley ...) so that the height of the peak is 5 mm, and about 2 m from the top (16) of this peak.
The m-shaped projection (68) is formed by folding back the part m at an angle (α) of 90 degrees or more so as to project inwardly from the peripheral edge, and the lining according to the third embodiment is formed. A base material (64) was produced. The backing substrate (6
In the base material body (66) of 4), there are elliptical liquid passage holes (70) having a width of about 1 mm and a length of about 10 mm.
18) at a 10 mm interval in each of the longitudinal direction and the transverse direction so that the longitudinal direction of 0) is the longitudinal direction of the substrate.
It was provided individually.

Then, in the same manner as in Example 2, the cultured epidermal cell sheet (22) prepared in the culture flask (20) was used.
2m after removing the culture medium and washing with phosphate buffer
Add L dispase solution (28) and add 25-37 ° C.
The culture flask (20) was heated so that the peripheral surface of the cultured epidermal cell sheet (22) was partially peeled off, and the backing substrate (64) according to Example 3 was subjected to chevron protrusions ( 6
8) It was placed in the center of the cultured epidermal cell sheet (22) so that the protruding direction of (folded portion) was upward. Then, when the cultured epidermal cell sheet (22) is peeled off from the bottom surface (21) of the culture flask (20) and contracted, the peripheral portion of the cultured epidermal cell sheet (22) is backed with tweezers to form a backing substrate (64). The cultured epidermal cell sheet (2
2) lined.

Then, the lined cultured epidermal cell sheet (22) is inverted in the culture flask (20) and left still so that the basal cell layer surface of the cultured epidermal cell sheet (22) faces upward. After removing the dispase solution (28), the operation of injecting and discharging the phosphate buffer was repeated several times to wash the cultured epidermal cell sheet (22) in the culture flask (20).

On the other hand, the blister case (72) having the brim (80) around the opening has a thickness of 5 in the dry state.
mm urethane foam (82) (manufactured by INOAC)
Was contained as a supporting substrate (78). Further, as shown in FIG. 9, the urethane foam (82) has a substantially rectangular recess (8) having an area slightly larger than the plane area of the lined cultured epidermal cell sheet (22), as shown in FIG.
4) (depth: about 1 mm) is provided, and a recess (8) for facilitating removal of the cultured epidermal cell sheet (22) is provided at the center of the four side walls of the recess (84).
6) (depth: about 1.5 mm) was provided.

Then, such a urethane foam (82)
The cultured epidermal cell sheet (22) backed is placed in the recess (84) so that the cultured epidermal cell sheet (22) is above the backing substrate (64) and does not contain serum. Replace the green medium (50) with urethane foam (8
In addition to the amount of filling the recess (84) of 2), the film-shaped lid body (76) was thermocompression-bonded to the brim portion (80) of the blister case (72) for packaging.

The above operation was carried out in a room temperature environment of about 19 to 26 ° C., and the packaged cultured epidermal cell sheet (22) was kept at a temperature environment of 2 to 9 ° C. for 24 hours. Was stored in a car-mounted electric refrigerator (about 5 to 8 ° C.) mounted on a sponge-like cushioning material having a thickness of 5 cm and transported to a medical institution for about 2 hours. Then, before being used in a medical institution, it was placed in a warmer at about 35 ° C. and then opened. The cultured epidermal cell sheet (22) in the liquid-tight container (72) had the same size as the backing substrate (64) and had a flat film-like shape on the backing substrate (64).

Example 4 As shown in FIG. 10, 22 acrylic pin-shaped protrusions ( 94) (tip diameter: about 1 mm, column diameter: about 0.8 mm, height: about 1.5 mm) is fixed vertically to the base material (92),
A backing substrate (90) according to Example 4 was produced. Also,
The acrylic sheet (92) has the liquid passage holes (94) with a diameter of about 3 mm at intervals of about 10 mm in a zigzag pattern, for a total of 1 hole.
Eight were provided.

Then, in the same manner as in Example 2 above, the cultured epidermal cell sheet (22) prepared in the culture flask (20) was used.
2m after removing the culture medium and washing with physiological saline
Add L dispase solution (28) and add 25-37 ° C.
The culture flask (20) was heated so that the peripheral edge of the cultured epidermal cell sheet (22) was partially peeled off, and the backing substrate (90) according to Example 4 was changed to a pin-shaped protrusion ( It was placed in the center of the cultured epidermal cell sheet (22) so that the protruding direction of 94) was upward. After that, when the cultured epidermal cell sheet (22) is peeled off from the bottom surface (21) of the culture flask (20) and contracted, the peripheral portion of the cultured epidermal cell sheet (22) is tipped with a backing substrate (90). The cultured epidermal cell sheet (22) was lined by hooking onto the pin-shaped protrusions (94) and leaving them in place.

Then, after removing the dispase solution (28) in the culture flask (20), the operation of injecting and discharging the phosphate buffer solution is repeated several times, so that the cultured epidermal cells are cultured in the culture flask (20). The sheet (22) was washed.

On the other hand, as shown in FIG. 12, the lid (3
4) and the container body (32) contained a water-absorbing polyester non-woven fabric (98) having a thickness of about 3 mm in a liquid-tight container (38) which was liquid-tightly sealed. Further, as shown in FIG. 12, the upper part of the non-woven fabric (98) is
A substantially rectangular recess having a slightly larger area than the planar area of the lined cultured epidermal cell sheet (22) (depth:
About 1 mm), and the cultured epidermal cell sheet (22) is provided in the central portion of the four side walls of the recess.
Recess for easy removal (depth: approx. 1.5 m
m) was provided.

Then, an amount of serum-free green medium (50) that fills the recess of the non-woven fabric (98) is added, and the recess of the non-woven fabric (98) is lined with the cultured epidermal cell sheet. (22) was placed such that the cultured epidermal cell sheet (22) was located above the backing substrate (90).

Further, on the cultured epidermal cell sheet (22) housed in the liquid tight container (38) as described above, the upper surface of the cultured epidermal cell sheet (22) and the liquid when the lid is closed. Space between the inner surface of the lid (39) and the inner surface of the lid (39): a substantially lattice-like silicone pressing member (100) having a thickness approximately the same as d is arranged, and the lid (34) The pressing member (1
At 00), the cultured epidermal cell sheet (22) was held while being pressed and packaged.

The above operation was carried out in a room temperature environment of about 19 to 26 ° C., and the packaged cultured epidermal cell sheet (22) was kept at a temperature environment of 2 to 9 ° C. for 24 hours. Were stored in a vehicle-mounted electric refrigerator (about 5 to 8 ° C.) mounted on a sponge-like cushioning material having a thickness of 5 cm and transported to a medical institution for about 2 hours. And before using it at a medical institution, after putting it in a warmer at about 35 ℃,
It was opened. The cultured epidermal cell sheet (22) in the liquid-tight container (38) had the same size as the backing substrate (90) and retained a flat film-like shape on the backing substrate (90).

Example 5 Cultured epidermal cell sheet (2 prepared in the same manner as in Example 2
2) is lined with the backing base material (90) according to Example 4 above, and then two sheets of medical gauze [capine: manufactured by Kawamoto Sangyo Co., Ltd.] folded in three are housed in a stack. In addition, the cultured epidermal cell sheet (22) lined with the backing base material (90) according to Example 4 is placed in a tight box structure container, and the cultured epidermal cell sheet (22) is more than the backing base material (90). It was placed on the upper side. afterwards,
After the serum-free green medium (50) is added to such an extent that the cultured epidermal cell sheet (22) is immersed to cover the lid, the clasps integrally provided on both end sides of the lid are attached to the container body. Locked to the engaging part that is integrally provided on the outer wall of
It was liquid-tightly sealed and packaged.

The above operation was carried out in a room temperature environment of about 19 to 26 ° C., and the packaged cultured epidermal cell sheet (22) was allowed to stand for 2 hours and then left for 24 hours. Stored in a temperature environment of 2-9 ° C,
It was transported to a medical institution in a vehicle-mounted electric refrigerator (about 5 to 8 ° C.) mounted on a sponge-like cushioning material having a thickness of 5 cm for about 2 hours. And before use in medical institutions, about 35 ℃
After being placed in a warmer, it was opened. The cultured epidermal cell sheet (22) in the liquid-tightly sealed container had the same size as the backing substrate (90) and had a flat film-like shape on the backing substrate (90).

Example 6 Rabbit cornea was separated into epithelium and parenchymal layer with tweezers and sterilized, and then the epithelial layer was cut with scissors and cut into 60 mm pieces.
It was attached to a Petri dish (Falcon) and the medium was added.
The medium is replaced every 3 days, and the cells are cultured for 6 days. When the epithelial cells proliferate from the fragmented portion, the epithelial cells are collected with trypsin, centrifuged, and the medium is added to the epithelial cell suspension. Was prepared. This was seeded in a 6-well dish (Falcon), the medium was replaced every 3 days, and the cells were cultured and proliferated to prepare sheet-like cultured corneal epithelial cells having a diameter of about 6 cm. The dispase solution was added here, and it left still at room temperature (about 23 degreeC).

On the other hand, diameter: 2.5 cm, thickness: about 0.1
In the same manner as in Example 3, the peripheral portion of the polyethylene disc film of mm was continuously processed so that the peaks (peaks-valleys) had a peak vertex pitch of 4 mm and a peak height of 4 mm. Then, about 2 mm from the apex of this mountain portion is bent inward at an angle of 90 degrees or more so as to project upward, that is, inclining inwardly from the peripheral edge portion to form a mountain-shaped protrusion. A backing substrate according to Example 6 was produced.

Then, when a part of the cultured corneal epithelial cell sheet was peeled from the well, the backing substrate according to Example 6 was placed in the center of the cultured corneal epithelial cell sheet and peeled from the well. The cultured corneal epithelial cell sheet was lined by hooking the peripheral edge of the cultured corneal epithelial cell sheet onto the chevron projections of the lining substrate with tweezers and leaving it in place. Then, the dispase solution in the wells was removed, and the lined cultured corneal epithelial cell sheet was repeatedly washed with physiological saline.

Then, the thus washed cultured corneal epithelial cell sheet is taken out from the well together with the backing substrate, and the bottomed cylindrical lid screw-engaged case structure container (diameter: 3.0 cm, depth : 0.7 cm) having a thickness of about 2 mm and having a backing base material on a water-absorbent polyvinyl alcohol sheet which is a supporting base material so that the application surface of the cultured corneal epithelial cell sheet is the upper surface. It was left still with the attached surface down. In addition, physiological saline in such an amount that the above-mentioned cultured epidermal cell sheet (22) was immersed was previously put in such a container.

Further, the cultured corneal epithelial cell sheet is covered with a polytetrafluoroethylene sheet having a thickness of about 2 mm as a pressing member in order to improve the stability during transportation, and then the opening of the container body is covered. The container was closed by screwing together the screw parts provided on the container body and the lid with the sealing member, the packaging was completed, and the container was transported to the animal house.

Then, when the sheet was made to open in the operating room of the animal house and the polytetrafluoroethylene sheet serving as the pressing member was removed, the cultured corneal epithelial cell sheet was formed into a flat film-like shape on the backing substrate at the size of the backing substrate. Was holding.

Then, in order to transplant the above-mentioned cultured corneal epithelium into a rabbit eye which has been peeled to a substantial part, the above-mentioned cultured corneal epithelial cell sheet is trimmed with experimental scissors together with the lining substrate immediately before the transplantation. And transplanted. Usually, when trimming, a cultured corneal epithelial cell sheet alone or a cultured corneal epithelial cell sheet lined with a conventional backing substrate is not stable, and it is difficult to trim to a desired shape. The above-mentioned cultured corneal epithelial cell sheet lined with the backing substrate according to the present example could be easily formed into a desired shape and had good operability.

Comparative Example 1 The cultured epidermal cell sheet (22) (about 10 cm × about 8 c) prepared in the culture flask (20) as in Example 2 above.
After removing the medium of m) and washing with a phosphate buffer,
Add 2 mL of dispase solution (28) and add 25-3
When the culture flask (20) was heated to 7 ° C. and a part of the periphery of the cultured epidermal cell sheet (22) was peeled off, no locking means was provided as a backing substrate, 9 cm. × 6 cm non-woven fabric made of chitin (Vesquitin W:
Using Unitika Ltd., the cultured epidermal cell sheet (22) was taken out from the culture flask (20).
In addition, packaging was performed in the same manner as in Example 2,
Used for storage / transportation.

When this was opened in a medical institution, the cultured epidermal cell sheet (22) showed that the medium (5
Due to 0), the non-woven fabric made of chitin was misaligned, about half of it was overlapped, and part of it was damaged, and it was impossible to secure the size at the time of packaging without retaining the flat film shape. .

Comparative Example 2 As in Example 2, the cultured epidermal cell sheet (22) (about 10 cm × about 8 c) prepared in the culture flask (20) was used.
After removing the medium of m) and washing with a phosphate buffer,
Add 2 mL of dispase solution (28) and add 25-3
When the culture flask (20) is heated to 7 ° C. and a part of the peripheral edge of the cultured epidermal cell sheet (22) is peeled off, no locking means is provided as a backing substrate, 8 The cultured epidermal cell sheet (22) was taken out from the culture flask (20) using a non-woven fabric made of chitin (Vesquitin W: Unitika Ltd.) having a size of 0.5 cm × 6 cm.

On the other hand, according to the method described in JP-A-8-23968, first, DME + 5% serum medium 100 m
1 was kept at 37 ° C., and bovine bone-derived gelatin powder 1
0 g was added, stirred and dissolved to prepare a 10% gelatin sol solution.

The cultured epidermal cell sheet (22) lined with the backing substrate as described above was placed in the center of a square Petri dish (manufactured by Nunc), and 20 ml of the gelatin sol described above was added around this. At room temperature (about 20
When left to stand at (° C.), it gelled after about 30 minutes, and the cultured epidermal cell sheet (22) was fixed by jelly-like highly viscous gelatin. This was further packaged in a container having a tight box structure and stored / transported.

Then, when opened in a medical institution, it was very difficult to take out the cultured epidermal cell sheet (22), and the taken out cultured epidermal cell sheet (22)
Was broken and could not retain the flat film shape.

Comparative Example 3 Cultured epidermal cell sheet (22) (about 10 cm × about 8 c) prepared in a culture flask (20) as in Example 2 above.
After removing the medium of m) and washing with a phosphate buffer,
Add 2 mL of dispase solution (28) and add 25-3
When the culture flask (20) is heated to 7 ° C. and a part of the peripheral edge of the cultured epidermal cell sheet (22) is peeled off, no locking means is provided as a backing substrate, S: Using 8 cm x 6 cm wide petrolatum gauze, remove the cultured epidermal cell sheet (22) from the culture flask (20), and further clip the petrolatum gauze and the cultured epidermal cell sheet (22) with a medical clip. With the lining cell base facing down so that the basal cell layer is on top, and this is placed in a blister case (72) having a brim (80) around the opening (length: about 100 mm x width: about 90 mm).
× depth: about 15 mm). Next, the phosphate buffer solution was repeatedly injected and discharged to wash the cultured epidermal cell sheet (22), and then the green medium (50) was added. After that, the film-shaped lid body (76) is hung on the opening of the blister case (72) and thermocompression-bonded with the brim case (80) of the blister case (72) to thereby form the blister case (72). The opening was closed in a liquid-tight manner, and it was transported to a medical institution in the same manner as in Example 2.

When this was opened at a medical institution and the cultured epidermal cell sheet (22) was taken out, the cultured epidermal cell sheet (22) was found to be clipped at the clipped portion of the cultured epidermal cell sheet (22) and petrolatum gauze. It was damaged. When the cultured epidermal cell sheet (22) is taken out, the central part is damaged and missing, and part of the end part is also damaged, and the flat membrane shape is not retained, and the damaged part is spread. Took a lot of trouble.

Then, in handling the membrane structure in Examples 1 to 6 and Comparative Examples 1 to 3 as described above,
Ease of taking out and the state of the film structure after taking out were evaluated, and the results are also shown in Table 1 below. The ease of removal was evaluated according to the following evaluation criteria.

-Easy to take-out-Good: Can be easily handled with fingers wearing tweezers or sterile gloves, and storage / transportation liquid does not spill. Almost good:
It can be easily handled with the hands of tweezers or sterile gloves, but care must be taken not to peel the membrane tissue from the backing substrate. Slightly poor: It cannot be easily handled with fingers wearing tweezers or sterile gloves, and some work such as re-expanding the membrane tissue is required. Poor: The membrane structure could not be taken out, or it could be taken out, but the flat film-like shape was not retained and it was damaged.

Further, in order to examine the biological activity of the membrane tissue, the whole of the membrane tissue taken out, or if not possible, part of it was made into single cells with trypsin, and the viable cell rate (viability) of the membrane tissue was measured. Was measured by the trypan blue dye exclusion method, and the results are also shown in Table 1 below.

[0147]

[Table 1]

As is clear from Table 1, in the membrane tissues according to Examples 1 to 6 stored / transported according to the present invention, the flat membrane-like shape of the membrane tissues can be advantageously retained,
Also, it can be seen that the handling is easy. Further, since the flat membrane-like shape of the membrane tissue is advantageously maintained, it is recognized that the biological activity of the membrane tissue is also sufficiently highly maintained.

On the other hand, in the case of the membrane structures of Comparative Examples 1 to 3, the operation of taking out from the container is complicated, and
It is recognized that the taken out membrane tissue does not remain flat membrane-like shape and the survival rate is lower than that of the membrane tissues according to Examples 1 to 6 above.

[0150]

As is apparent from the above description, in the backing base material for a membrane tissue according to the present invention, since the locking means is provided at the peripheral portion thereof, the membrane structure is provided in the locking means. The peripheral edge portion of the backing is hooked and locked so that the membrane structure can be advantageously retained on the backing base material. Therefore, the membrane structure is prevented from peeling or detaching from the backing base material. That is very effectively avoided.

Further, in the method for storing / transporting a membrane structure according to the present invention, since the above-mentioned backing base material for a membrane structure is used, the membrane structure is peeled or detached from the backing base material. Can be prevented very effectively and, therefore, such membranes are preserved in the preservation / transport solutions used to preserve the biological activity of the membrane tissue during normal storage and transport steps. It is possible to effectively prevent the occurrence of problems such as the tissue floating, breaking, or twisting, which makes it impossible to maintain the flat membrane shape or damage. .

Moreover, the storage / transportation liquid is housed together with the lined membrane tissue in the liquid-tight container so that the storage / transportation liquid can always keep the membrane tissue in a wet state. Therefore, the biological activity can be favorably retained without impairing the biological properties of the membrane tissue.

[Brief description of drawings]

FIG. 1 is an explanatory view showing a specific example of a backing base material for a membrane structure according to the present invention, in which (a) is a plan explanatory view,
(B) is a side view, and (c) is a cross-sectional view taken along the line I-I in (a).

FIG. 2 is an explanatory view showing a step of lining a membrane structure with the backing substrate shown in FIG. 1, and FIG. 2 (a) is a vertical cross-sectional explanatory view showing a state in which the backing substrate is arranged on the membrane structure. And
(B) is a longitudinal cross-sectional explanatory view showing a state in which the peripheral portion of the membrane tissue is locked by the locking means of the backing substrate.

3 is an explanatory plan view of a film structure lined with the backing substrate shown in FIG. 1. FIG.

FIG. 4 is a longitudinal cross-sectional explanatory view showing a state in which the membrane tissue lined with the backing base material shown in FIG. 1 is arranged on a supporting base material in a liquid-tight container in which a storage / transport liquid is enclosed. Is.

FIG. 5 is an explanatory view showing a step of preparing the supporting base material shown in FIG. 4 and filling it with a storage / transport liquid,
FIG. 3 is a vertical cross-sectional explanatory view showing a step of flowing an agar sol into a liquid-tight container, and (b) shows a step of injecting a storage / transport solution into an agar gel produced by solidifying the agar sol. FIG.

FIG. 6 is an explanatory view showing another specific example of the backing base material for a membrane structure according to the present invention, in which (a) is a plan explanatory view,
(B) is a side view, and (c) is a VI-VI partial enlarged sectional view of (a).

FIG. 7 is an explanatory view showing a step of lining the film structure with the backing base material shown in FIG. 6, and FIG. 7A is an end face explanatory view showing a state where the backing base material is arranged on the film structure. Yes,
(B) is an end face explanatory view showing a state in which the peripheral edge portion of the membrane tissue is locked by the locking means of the backing base material.

8 is a cross-sectional explanatory view showing a state in which the membrane tissue lined with the backing base material shown in FIG. 6 is arranged on a supporting base material in a liquid-tight container in which a storage / transport liquid is enclosed. is there.

9A and 9B are explanatory views showing a housed state of the supporting base material shown in FIG. 8, FIG. 9A is a plan explanatory view, and FIG. 9B is an IX-IX sectional explanatory view in FIG. 9A. .

FIG. 10 is an explanatory view showing still another specific example of the backing substrate for a membrane tissue according to the present invention, (a) is a plan explanatory view, (b) is a side explanatory view, c) is (a)
FIG. 6 is an enlarged cross-sectional explanatory view taken along line XX in FIG.

FIG. 11 is an explanatory view showing a step of lining the membrane structure with the backing base material shown in FIG. 10, wherein (a) is a longitudinal cross-sectional explanatory view showing a state in which the backing base material is arranged on the membrane structure. FIG. 3B is a vertical cross-sectional explanatory view showing a state in which the peripheral edge portion of the membrane tissue is locked by the locking means of the backing base material.

FIG. 12: The membrane tissue lined with the backing base material shown in FIG. 10 is placed on the support base material in the liquid-tight container in which the storage / transport liquid is sealed, and immovable by the holding member. FIG. 4A is an explanatory view showing a state in which the
It is a top surface explanatory view, (b) is XII-XII in (a).
FIG.

[Explanation of symbols]

10, 64, 90 Backing base material 12, 66, 92
Base material main body 14,68 Angle projection 18,70,96
Liquid passage hole 19,20 Culture flask 22 Membrane tissue 32,74 Container body 34,76 Lid 36 Seal member 38,72 Liquid-tight container 50 Storage / transport liquid 52,78,98
Support substrate 94 Pin-shaped protrusion 100 Holding member

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) // (C12N 1/04 C12N 5/00 E C12R 1:91) (72) Inventor Nobutaka Yamamoto Aichi Prefecture 5-10 Takamoridai, Kasugai-shi, 10-shares in the Menicon Research Institute, Inc. (72) Inventor, Hisae Hayashi 5--10-1, Takamoridai, Kasugai-shi, Aichi Prefecture, F-Term (Reference) 3E067 AB99 AC01 BA01A EE26 GA21 4B029 AA21 BB11 CC02 CC11 4B065 AA93X BD12 CA44 4C081 AC16 BB01 BC02 CA051 CA161 CA231 CB011 CC01 CD011 CD121 CD151 DA01 DA02 DB07 DC04 DC05

Claims (20)

[Claims] 1. A backing base material having a flat plate shape as a whole, which is arranged on one surface side of a flat membrane-like membrane tissue made of a bio-derived material to hold the membrane tissue, A backing base material for a membrane tissue, wherein locking means for hooking and locking at least a peripheral edge portion of the membrane tissue is provided on at least a peripheral edge portion of the base material. 2. The backing substrate for a membrane structure according to claim 1, which has a plane area smaller than a plane area of the membrane structure. 3. The locking means is provided on at least a part of a peripheral edge of a surface of the base material on the side facing the film structure, inclined outward from the peripheral edge or with respect to the base material surface. The membrane according to claim 1 or 2, which is provided so as to protrude in a vertical direction, and the peripheral edge portion of the membrane tissue is retained by the retaining means to retain the membrane tissue. Tissue backing substrate. 4. The locking means is provided on at least a part of the peripheral edge of the base material so as to project outward from the peripheral edge and in a direction parallel to the surface of the base material. The backing substrate for a membrane tissue according to claim 1 or 2, wherein the membrane tissue is held in a state of being in contact with one surface of the membrane tissue by locking the peripheral edge portion of the membrane tissue. Material. 5. The locking means is provided on at least a part of a peripheral edge of a surface of the base material opposite to a surface facing the film structure, and is inclined inward from the peripheral edge. It is provided so as to project in a direction perpendicular to the surface of the base material, and is brought into contact with one surface of the film tissue by locking the locking means so as to wrap it around the peripheral edge of the film tissue. The backing substrate for a membrane tissue according to claim 1 or 2, wherein the membrane tissue is retained in the state. 6. The backing substrate for a membrane tissue according to claim 1, wherein a liquid passage hole that allows a liquid to pass therethrough is provided so as to penetrate the substrate. 7. The backing substrate for a membrane tissue according to claim 1, which is in the form of a sheet, a woven fabric, a non-woven fabric, or a sponge made of a bio-derived polymer or a synthetic polymer. Material. 8. A method for storing or transporting the membrane tissue using the backing substrate according to any one of claims 1 to 7, wherein the membrane tissue is held in the locking means of the backing substrate. The peripheral portion is locked, and the backing base material is fixedly arranged on one surface side of the membrane tissue, so that the membrane tissue is lined, and the lined membrane tissue is fixed with a predetermined liquid. Characterized in that it is housed in a tight container and is stored or transported while the membrane tissue is maintained in a sufficiently wet state with a storage / transportation liquid and the flat membrane shape of the membrane tissue is maintained. A method for storing / transporting a membrane tissue. 9. The film structure according to claim 8, wherein the film structure backed by the backing base material is accommodated so that the film structure is located above the backing base material. How to store / transport. 10. The membrane structure backed by the backing base material is placed on a hydrophilic or water-absorbing support base material and housed in the predetermined liquid-tight container. The method for preserving / transporting a membrane tissue according to claim 8 or 9. 11. The backing base material is further housed in the liquid-tight container, and the backing base material is pressed by the holding member to press the membrane tissue backed by the backing base material. The method for preserving / transporting a membrane tissue according to any one of claims 8 to 10, wherein the membrane tissue lined with (4) is fixedly held in the liquid-tight container. 12. The pressing member has a thickness corresponding to a space between the lined membrane structure placed on the supporting base material and the inner surface of the liquid tight container facing the lined film structure. Item 12. The method for preserving / transporting a membrane tissue according to Item 11. 13. The preservation of the membrane tissue according to claim 8, wherein the contraction action of the membrane tissue causes the peripheral portion of the membrane tissue to be locked by the locking means of the backing substrate. /
Transportation method. 14. The supporting substrate according to claim 8, which is in the form of a sheet, a woven fabric, a nonwoven fabric, or a sponge made of a bio-derived polymer or a synthetic polymer. Method for preserving / transporting membrane tissue of the above. 15. The method for preserving / transporting a membrane tissue according to claim 8, wherein the membrane tissue is a biological membrane tissue. 16. The method for preserving / transporting a membrane tissue according to claim 8, wherein the membrane tissue is cultured skin or a cultured cornea. 17. The method for preserving / transporting a membrane tissue according to claim 8, wherein the membrane tissue is a cell sheet. 18. The method for preserving / transporting a membrane tissue according to claim 8, wherein the membrane tissue is a cultured epidermal sheet, a cultured mucous membrane sheet or a cultured corneal epithelial sheet. 19. The method for preserving / transporting a membrane tissue according to claim 8, wherein the storage / transportation liquid is a liquid capable of maintaining the physiological activity of the membrane tissue. 20. The liquid-tight container is a blister case,
The method for preserving / transporting a membrane tissue according to any one of claims 8 to 19, which is a tight box or a lid screw-engagement type case.