KR20040050711A - Cultured cell seat packing member and seat inserting member - Google Patents

Abstract

PURPOSE: Provided are a cultured cell sheet packing member and a sheet inserting member which prevent the cultured cell sheet from going wrong, being damaged or being flamed when transported. CONSTITUTION: The cultured cell sheet packing member includes: a tray(21), made out of an air- and liquid-tight material; a lid(31), made out of an air- and liquid-tight material; a liquid medium, filled into a sealed space which is covered with the tray(21) and the lid(31), and has a complete vacuum state; and a cultured cell sheet, immersed into the liquid medium inside the sealed space; besides a first interposition member, arranged between the cultured cell sheet and the tray(21) and contacting with the tray(21), but partially with the cultured cell sheet; and a second interposition member, arranged between the cultured cell sheet and the lid(31), and contacting with the lid(31), but partially with the cultured cell sheet.

Description

CULTURED CELL SEAT PACKING MEMBER AND SEAT INSERTING MEMBER}

The present invention relates to a culture cell sheet package and a sheet fitting member.

In recent years, attention has been paid to tissue engineering and regenerative medicine for treatment by incubating cells and tissues in vitro (out-of-living) and applying them to living bodies again. Particularly, a technique of applying a part of the patient's or donor's skin or oral mucosa and the like to be cultured in a sheet form and then applied to a living body has been put into practical use. For example, in the case of the skin, treatment of birthmarks, ulcers, lacerations, tattoos, and the like, and transplantation of cultured epidermal (epidermal) cell sheets on skin extracting skins for transplantation have been attempted. In the case of oral mucosa, transplantation of cultured cell sheets using oral mucosal epithelial cells has been attempted for the treatment of wounds and defects of gums and mucosal tissues caused by oral diseases such as periodontal disease and malignant tumors. These cultured cell sheets are prepared using a technique of tissue engineering, cultured in a culture flask so that epithelial (epidermal) cells have a thin sheet-like structure in which 5 to 10 layers are layered, and then separated from the culture flasks for transplantation. Is being provided. However, the production of such cultured cell sheets is not complicated and can be easily performed in an ordinary medical institution. Therefore, the transplantation treatment of these culture cells is only carried out at specialized facilities such as university hospitals with sufficient culture facilities and skilled culture techniques. In addition, in the case of transport by a culture flask, the peeling operation of the culture cell sheet is performed in a medical institution, so additional work is required before the operation, which burdens the operator.

In this regard, if a culture cell sheet produced in a specialized facility is properly packaged and can be transported stably, it can be easily applied in most medical institutions having a general surgical facility, and many patients may not receive the transplantation treatment of cultured cells. Can be. As this type of cultured cell sheet package, a cultured epithelial cell sheet clipped to a substrate sheet is mounted on the bottom of a tray recess, for example, as disclosed in Japanese Patent Publication No. 2733800. It is known that the tray was sealed by a sheet-like lead after the insertion of the mold.

However, in the cultured cell sheet package of Japanese Patent No. 2733800, there is a fear that air remains inside during packaging. If air remains inside, the culture medium may sway due to shaking during transportation or bubbles may move in the culture medium, causing the cultured epithelial cell sheet to shift or become defective. There is a concern. In addition, since the gap between the cultured epithelial cell sheet and the lead clipped to the substrate sheet is empty, if the cultured cell sheet package is shaken during transportation, the cultured epithelial cell sheet and the lead may collide and the cultured epithelial cell sheet may be damaged. .

An object of the present invention is to provide a culture cell sheet package which prevents the culture cell sheet from being misaligned, missing or degraded during transportation.

1 is a manufacturing flow of a cell sheet-containing carrier.

2 is an assembled perspective view of the culture cell sheet package.

3 is a cross-sectional view (left half) of the cultured cell sheet package.

4 is an assembled perspective view of the culture cell sheet package.

5 is a cross-sectional view (left half) of the culture cell sheet package.

6 is an enlarged view of a fitting portion of the net.

* Explanation of symbols for the main parts of the drawings *

10: cultured epithelial cell sheet

12: culture flask

14: first carrier

16: second carrier

18: cell sheet-containing carrier

20: Packing Container

21: tray

22: tray recess

23: tray embankment

24: tray groove

25: tray collar

26: annular step

27: triangular step

31: lead

32: lead recess

33: lead collar part

33: Reed Dike

34: lead groove

35: lead collar part

41: seal

45,46: Net

50: cultured cell sheet package

The cultured cell sheet package of the present invention adopts the following means to achieve the object described above.

That is, the cultured cell sheet package of the present invention,

A tray made of airtight and dense material,

A lid made of airtight and dense material,

A liquid medium filled with air almost completely removed from the sealed space surrounded by the tray and the lid;

It is provided with a culture cell sheet immersed in the liquid medium in the sealed space.

In this culture cell sheet package, since the liquid medium filled in the sealed space is almost completely removed from the air, the culture cell sheet immersed in the liquid medium is not deteriorated by air. In addition, even if the cultured cell sheet package is shaken during transportation, there is no fear that the cultured epithelial cell sheet may be displaced or missing since the liquid medium does not stray or bubbles are not moved during the liquid medium. In addition, in "liquid medium filled with air almost completely removed", "air almost completely removed" means that air is removed to such an extent that these effects are exhibited. It also includes a few remaining cases.

Cultured cell sheet package of the present invention,

A first interposition member between the culture cell sheet and the tray and in contact with the tray and at least partially in contact with the culture cell sheet;

A second interposition member may be provided between the culture cell sheet and the lid and in contact with the lead and at least partially in contact with the culture cell sheet.

In this cultured cell sheet package, the cultured cell sheet is supported in the liquid medium by the first interposition member and the second interposition member. For this reason, even if the culture cell sheet package is shaken at the time of transport, the culture cell sheet does not move easily, and there is no fear that the culture cell sheet collides with the lid or the tray and is damaged. In addition, since the culture cell sheet and the first and second interposition members are only partially in contact with each other rather than being in full contact, the culture cell sheet is hardly damaged by the first and second interposition members. . Therefore, according to the cultured cell sheet package, the cultured cell sheet can be transported without damaging the tray, the lid and the cultured cell sheet.

In the present invention, the term "cultured cell sheet" refers to a cultured epidermal (epidermal) cell sheet of monolayer or plural layers (layered) consisting only of cells as described in the [Prior Art] column. It includes the thing cultured on the sheet | seat scaffold comprised from a gel, a fibrin gel, etc. In addition, sheet-like cell insertion type such as cultured dermis seeded and cultured with fibroblasts with a porous body such as collagen sponge or hyaluronic acid sponge, or complex cultured skin having epithelial (epidermal) cell layer formed on the cultured dermis. Include implants.

The material of the "tray", "lead", "first intervening member" and "second intervening member" is not particularly limited as long as it is inert to the liquid medium or culture cell sheet. For example, polyethylene, polypropylene, polystyrene, polycarbonate And synthetic resins such as polyvinyl chloride and Teflon (registered trademark), hydroxyapatite ceramics, alumina ceramics, glass and the like are preferably used. The "first interposition member" and "second interposition member" preferably have flexibility that does not cause damage to the culture cell sheet. For example, an elastic material such as silicone rubber is used in addition to the synthetic resin described above.

In the culture cell sheet package of the present invention, the culture cell sheet may be in contact with the first interposition member and the second interposition member in a state of being supported by a flat support. In this case, the cultured cell sheet is reinforced by a flat support, which can prevent the cultured cell sheet from being damaged during handling even when the cultured cell sheet does not have sufficient strength. Here, in supporting a culture cell sheet on a flat support, one support side and a culture cell sheet may be superimposed, one support may be packaged in a culture cell sheet, and one support may be folded and cultured therebetween. The cell sheet may be packaged, a cultured cell sheet may be sandwiched between two supports, or any other method may be adopted.

Thus, when using the culture cell sheet supported by the support body, the said support body may be formed from the wound coating material. The support formed of the wound coating material is suitable for supporting the culture cell sheet, and also has good workability when releasing the support of the culture cell sheet. Here, as a wound coating material, For example, biological component coating materials, such as a sterile gauze and Beskichin (trademark), Arginate coating materials, such as a sorbic acid (trademark), Hydrocolloid dressings, such as Duoactive (trademark), Bioclo Polyurethane film dressings, such as a sheave (trademark), etc. are mentioned. When such a wound coating material is used as a support for supporting one side of a culture cell sheet, the wound coating material can be applied as a wound coating material as it is after being applied to a transplantation site. The support may be formed of a flexible polymer material (eg, silicone rubber) that is not limited to these and does not damage the culture cell sheet. Further, collagen gels, fibrin gels, gelatin, agar media, etc. may be sheet-shaped, sterile paper, sterile nonwoven fabrics, or the like.

In addition, in the case of using the culture cell sheet supported by the support, the support is composed of a first support sheet and a second support sheet, the first support sheet is one side of the first support sheet overlaps with the culture cell sheet is integrally integrated. The second support sheet may be supported in a packaged state of the culture cell sheet integrated with the first support sheet. In this case, since the cultured cell sheet is reinforced by the first support, the cultured cell sheet can be prevented from being damaged during handling even when the cultured cell sheet does not have sufficient strength. In consideration of facilitating the handling at the time of transplantation, it is preferable to support the opposite side of the implantation application surface (surface in contact with the transplantation site) of the culture cell sheet with the first support sheet. In this case, the transplantation application surface of the culture cell sheet is not covered by the first support sheet but is protected by the packaging by the second support sheet. In particular, in the case of cultured epidermal (epidermal) cell sheets, it is possible to reliably protect the basal layer having the cell proliferation ability of the transplantation application surface. Moreover, it is preferable to form the opening part of a 2nd support sheet in a lead side. In this case, the culture cell sheet supported on the first sheet can be easily taken out while leaving the second support sheet in the tray.

In the culture cell sheet package of the present invention, the position at which the first interposition member contacts the culture cell sheet and the position at which the second interposition member contacts the culture cell sheet may not be opposed to each other. If both positions are opposite, the culture cell sheet may be caught at that position when the tray or lid is deformed, and excessive pressure may be applied.However, if both positions are not opposed and shifted, the culture cell sheet may be deformed even if the tray or lid is deformed. It is preferable because there is no fear of excessive pressure because it is not jammed.

In the cultured cell sheet package of the present invention, the first interposition member is formed separately from the tray or integrally molded with the tray, and the second interposition member is formed separately from the lead. Or may be molded integrally with the lead. In the case where the first interposition member or the second interposition member is integrally molded with the tray or the lid, there is no need to treat both the interposition members separately, so that the operability at the time of packaging or unpacking is excellent. On the other hand, when the first interposition member or the second interposition member is formed separately from the tray or lead, the effects of the present invention can be obtained by using the existing tray and lead. Specifically, (1) the first interposition member may be integrally molded with the tray, the second interposition member may be integrally molded with the lead, and (2) the first interposition member is integrally molded with the tray and the second interposition is formed. The member may be molded separately from the lid, ③ the first interposition member may be formed separately from the tray, the second interposition member may be integrally formed with the lid, and ④ the first interposition member may be formed from the tray. It may be molded separately and the second interposition member may be molded separately from the lead.

In the cultured cell sheet package of the present invention, the first interposition member may be a lattice-shaped wire member or a plurality of protrusion members, and the second interposition member may be a lattice-shaped wire member or a plurality of protrusion members. When the wire member is adopted, the contact with the culture cell sheet is approximately line contact, and when the protrusion member is adopted, the contact with the culture cell sheet is approximately point contact, so that the burden is not applied to the culture cell sheet either way. . Specifically, (1) both the first interposing member and the second interposing member may be a lattice-shaped wire member, (2) both the first interposing member and the second interposing member may be a plurality of protrusion members, and (3) the first interposing member is lattice-shaped. The first interposition member may be a plurality of protrusion members, the first interposition member may be a plurality of protrusion members, and the second interposition member may be a lattice-shaped wire member.

Here, the first interposition member is a net separate from the tray, or a plurality of protrusion members integrally formed with the tray, and the second interposition member is a net separate from the lead, or It may be a plurality of projection members integrally molded.

In the culture cell sheet package of the present invention, it is preferable that the first interposition member and the second interposition member are in contact with the culture cell sheet with a pressing force such that the culture cell sheet is not damaged by both members. In this way, it is possible to reliably prevent the cultured cell sheet packaged in the cultured cell sheet package from being damaged by both intervening members. For example, the height of the enclosed space minus the sum of the height of the first interposition member and the height of the second interposition member is approximately equal to the thickness of the culture cell sheet (including the support when supported by the support), You may make it slightly smaller than thickness.

The cultured cell sheet package of the present invention may be used for transporting the cultured cell sheet as a use. The cultured cell sheet should not be damaged when the cultured cell sheet is manufactured in a specialized facility and transported during the medical period. According to the cultured cell sheet package of the present invention, the cultured cell sheet in a peeled state is easy to handle in a medical field. Even if it is transported, the demand can be satisfied.

The cultured cell sheet package of the present invention includes a tray recess formed in the center of the tray and a lead recess formed in the center of the lid, and the sealed space surrounded by the tray and the lid includes the tray recess. And a space lying between the lead recesses. By doing so, it is possible to prevent the liquid medium filled in the sealed space from flowing out or overflowing from the tray when the lid is opened. In addition, in the cultured cell sheet package, it is not preferable that air remains in the sealed space. Therefore, air is required to be removed from the sealed space during packaging. It is possible to easily remove air from the enclosed space by injecting more into the tray than the filling amount and removing the air with a little liquid medium when sealing with the lid. In addition, it is preferable that each concave portion has a lower area than the culture cell sheet (area larger than the first support when the first support is used, and larger than the second support when the second support is used).

Thus, the cultured cell sheet package having a tray recess and a lead recess includes a tray bank part formed on an outer circumference of the tray recess, a tray groove part formed on an outer circumference of the tray bank part of the tray, and A lead embankment portion formed on an outer periphery of a lead recess, and a lead groove portion formed on an outer periphery of the lead embankment portion among the leads; The tray groove portion and the lead groove portion may be in close contact with each other. In this way, it is possible to reliably prevent the liquid medium filled in the sealed space from flowing out or overflowing from the tray when the lid is opened. In addition, air can be removed from the sealed space more easily.

In this case, a hole may be formed in at least one of the tray groove portion and the lead groove portion. In this case, a small amount of liquid medium and air removed from the sealed space at the time of sealing passes through the gap between the tray bank and the lead bank, and then is discharged to the outside from the hole formed in the tray groove or the lead groove. Air can be removed from the In addition, you may form the hole which has the same function also in a tray bank part and a lead bank part.

In the cultured cell sheet package of the present invention, the tray and the lid each have a tray collar portion and a lead collar portion, and the tray and the lid are attached to each other so as to cover a seal material of a different material from the tray and the lid. The lead may be sealed. In this way, when removing a sealing material, since a sealing material is a material different from a tray and a lid, it is easy to take out.

The sheet fitting member of the present invention is a sheet fitting member for use in a culture cell sheet package in which a culture cell sheet is immersed in a liquid medium filled in a state where air is almost completely removed from a sealed space surrounded by a tray and a lid. A first fitting member partially contacted with the culture cell sheet between the sheet and the tray and formed separately from the tray, partially contacting the culture cell sheet between the culture cell sheet and the lid, Is characterized in that it comprises a second fitting member formed separately and engaging means for engaging the first fitting member and the second fitting member substantially integrally. In this way, after opening the culture sheet package, the culture cell sheet securely supported by the first and second fitting members can be easily taken out. In addition, when the cultured cell sheet not packed with the support is sandwiched between the first and second fitting members, the cultured cell sheet can be easily washed in that state. On the other hand, the engaging means is preferably a means that can be easily separated after engaging the first fitting member and the second fitting member. For example, the convex portion and the concave portion may be formed in each fitting member as a fastening means so that the concave portion and the convex portion are engaged with each other when the culture cell sheet is fitted, and the recessed portion and the convex portion engaged with the culture cell sheet may be separated. . The gap between the first fitting member and the second fitting member when engaged by the engaging means is preferably designed to be equal to or slightly larger than the thickness of the culture cell sheet to be fitted or the thickness of the culture cell sheet and the support. . In addition, both members are arranged in a line so that one side of the first fitting member and one side of the second fitting member are in common, and the common side is folded with gold to fold the culture cell sheet (or the culture cell sheet and the support). You may insert it.

EXAMPLE

1. Preparation of cultured epithelial cell sheet

About 1 cm <2> of skin tissue was taken from the patient who performed an informed outlet. The collected skin tissue was separated into a dermal layer and an epidermal layer by treatment in a medium for epithelial detachment, and an epithelial cell suspension was prepared by trypsinizing the epidermal layer. Subsequently, feeder cells were placed on the bottom of the culture flask, and then the epithelial cell suspension was seeded. As feeder cells, mouse fibroblasts whose proliferative capacity was inhibited by mitomycin C were used. After cell seeding, 5v / v% FBS (fetal bovine serum) -DMEM (Dulbecco's modified eagle medium) was injected as a culture medium, and epithelial cells were cultured in the presence of feeder cells. After colony formation of epithelial cells was confirmed, the cultured epithelial cell sheets were formed by stratification (sheeting) through a confluent state by continuously culturing. By the time the cultured epithelial cell sheet was formed, most of the feeder cells were removed from the culture system.

2. Preparation of Cellular Sheet Carrier

The manufacturing method of a cell sheet containing carrier is demonstrated based on FIG. Peeling medium (400PU / mL dispase medium) was added to the culture flask 12 in which the cultured epithelial cell sheet 10 was incubated and incubated for 30 minutes in a CO ₂ incubator (37 ° C, 10% CO ₂ ). . Thereafter, when the culture flask 12 was a fill-off flask, the sealing member of the upper surface was peeled off, and in the case of a normal culture flask, the upper surface portion was cut with a hot knife. Subsequently, after confirming that a portion around the culture epithelial cell sheet 10 was peeled from the culture flask 12 using tweezers, the medium was taken out and washed twice with PBS (phosphate buffer). After washing, the tweezers were scraped off around the culture epithelial cell sheet 10, and then pulled out with a small amount of PBS (see Fig. 1 (a)). On the other hand, a sterile nonwoven fabric having an area slightly smaller than the total area of the cultured epithelial cell sheet 10 is prepared as the first carrier (first support) 14, and the first carrier 14 is placed in the culture flask 12. Overlaid on the cultured epithelial cell sheet 10 (see FIG. 1 (b)), the portion protruding from around the first carrier 14 of the cultured epithelial cell sheet 10 is folded back into the first carrier 14. The cultured epithelial cell sheet 10 and the first carrier 14 were integrated (see FIG. 1 (c)). In the culture flask 12, the cultured epithelial cell sheet 10 has a base layer, which is a transplantation application side facing downward, so that the first carrier 14 covers the side opposite to the implantation application surface. On the other hand, although not shown, a small amount of liquid medium (transport medium) is placed in a sterilization tray, and a sterile nonwoven fabric having an area of about 2.5 times the first carrier 14 is used as the second carrier (second support) 16. The second carrier 16 was prepared and laid in a sterile tray. Then, the cultured epithelial cell sheet 10 integrated with the first carrier 14 is suspended, mounted on the second carrier 16 in the sterilization tray so that the transplantation application side is faced up, and wrapped with the second carrier 16. (See FIG. 1 (d)) to obtain a cell sheet-containing carrier 18 (see FIG. 1 (e)). That is, the cell sheet-containing carrier 10 is a package in which the cultured epithelial cell sheet 18 is packaged with the first carrier 14 and the second carrier 16. In addition, the opening part of the 2nd carrier 16 at this time was provided upward.

3. Preparation of cultured cell sheet package-1

2 is an assembled perspective view of the culture cell sheet package. The packaging container 20 is composed of a tray 21, a lid 31, and a seal 41. The tray 21 is made of polypropylene and has a tray recess 22 capable of storing a liquid medium, a tray embankment 23 formed around the tray recess 22, and a tray embankment 23. The tray groove part 24 formed along the outer periphery of the tray groove part 24, and the tray collar part 25 formed along the outer periphery of this tray groove part 24 are provided. An annular step 26 is formed on the inner circumferential side of the tray collar 25, and a triangular step 27 is formed at one of the four corners of the tray collar 25. On the other hand, the lid 31 is made of polypropylene and has a lid recess 32 which is shallower than the depth of the tray recess 22, a lid embankment 33 formed around the lead recess 32, and The lead groove part 34 formed along the outer periphery of the lead bank part 33 and the lead collar part 35 formed along the outer periphery of this lead groove part 34 are provided. And the lead recessed part 32 is inserted into the tray recessed part 22, the tray bank 23 and the lead bank 33 are made to match, and the tray groove part 24 and the lead groove part 34 are matched, When the lead collar portion 35 is superimposed on the tray collar portion 25, the lead collar portion 35 is in a state coinciding with the annular terminal 26 of the tray collar portion 25, and the seal 41 is here. When the tray collar portion 25, the lead collar portion 35, and the seal 41 are thermally welded with each other, a sealed space S surrounded by the tray recess 22 and the lead recess 32 is formed. However, the seal 41 and the triangular step 27 are not thermally welded so that the seal 41 can be removed from the triangular step 27 when the seal 41 is removed. In addition, the material of the seal 41 is preferably a material different from the tray 21 and the lid 31. In this way, the seal 41 can be easily removed from the tray 21 and the lid 31 at the time of opening.

In addition, a transport medium (antibiotic-containing DMEM) is placed in the tray recess 22 of the tray 21, and the cell sheet-containing carrier 18 produced in the above 2. is loosened from the top and bottom with the nets 45 and 46. It was inserted and immersed in the medium for transport. As the nets 45 and 46, the material was polypropylene, and the wire member of thickness 2mm was enclosed longitudinally and horizontally at the predetermined space | interval (for example, 1 cm). The wire member has a thickness or a predetermined interval determined according to the size of the cultured cell sheet, and the like, and the thickness or the interval is determined so that the minimum contact area can reliably support the cultured epithelial cell sheet 10. It is preferable for suppressing damage to (10). Thereafter, the lead recesses 32 are inserted into the tray recesses 22, and the tray banks 23 and the lead banks 33 are aligned so that no air remains in the sealed space S, and the tray recesses are provided. When the lead groove portion 34 is aligned with the lead groove portion 34 and the lead collar portion 35 is superimposed on the tray collar portion 25, the lead collar portion 35 is the annular terminal 26 of the tray collar portion 25. And the seal 41 was covered, and the tray collar portion 25, the lead collar portion 35, and the seal 41 were thermally welded. As a result, the sealed space S enclosed by the tray recess 22 and the lead recess 32 is filled with a transport medium, and the cell sheet-containing carrier 18 is formed into the net 45, 46 in the transport medium. It became the state supported by the, and the cultured cell sheet package 50 was completed. The cross section (left half) of this culture cell sheet package 50 is shown in FIG. In addition, the above operation was performed in a clean room. The packaging containers 20 and the nets 45 and 46 were sterilized in advance.

In this cultured cell sheet package 50, a net 45 interposed between the cell sheet-containing carrier 18 and the bottom surface of the tray recess 22 is substantially in line contact with the cell sheet-containing carrier 18, and at the same time the tray. The net 46 which is also substantially in line contact with the bottom surface side of the recess 22 and which is interposed between the cell sheet-containing carrier 18 and the bottom of the lead recess 32 is approximately in the cell sheet-containing carrier 18. The cell sheet-containing carrier 18 is supported in the transport medium by being in linear contact and substantially linearly in contact with the rear face of the bottom face of the lead recess 32. In addition, the thickness of the net 45 and the thickness of the net 46 at a distance from the bottom surface side of the tray recess 22 to the bottom rear face side of the lid recess 32 (that is, the height of the sealed space S). Is subtracted from the thickness of the cell sheet-containing carrier 18 or slightly smaller than the thickness of the cell sheet-containing carrier 18. For this reason, the force by which the nets 45 and 46 pressurize the cell sheet containing carrier 18 is weak.

According to the culture cell sheet package 50 described in detail above, since the liquid medium filled in the sealed space S is almost completely removed, the culture cell sheet immersed in the liquid medium may be deteriorated by air. There is no. In addition, even if the culture cell sheet package 50 is shaken at the time of transport, the culture medium epithelial cell sheet does not have to be shifted or missed because the bubble does not move or the bubbles do not move during the liquid medium. 18) is not easily moved or released, and there is no fear that the cultured epithelial cell sheet 10 contained in the cell sheet-containing carrier 18 collides with the tray 21 or the lid 31 and is damaged. In addition, the cell sheet-containing carrier 18 and the nets 45 and 46 are not in full contact but merely in line contact, and the nets 45 and 46 pressurize the cell sheet-containing carrier 18. Since the force to perform is weak, the cultured epithelial cell sheet 10 contained in the cell sheet-containing carrier 18 is hardly damaged by the nets 45 and 46. Therefore, the cultured epithelial cell sheet 10 (particularly, the base layer important for engraftment and healing of the cultured epithelial cell sheet 10) can be transported without damage.

In the use, the sealed space S is formed by the recesses 22 and 32 of the tray 21 and the lid 31, so that the liquid medium filled in the sealed space S at the time of opening the lid 31 flows. It can prevent falling. In addition, after opening the lid 31 and removing the net 46 by opening the opening of the second carrier 16 constituting the cell sheet-containing carrier 18 to the lead side, the first carrier is opened by opening the second carrier 16. The cultured epithelial cell sheet 10 suspended in 14 is exposed and can be easily taken out.

In addition, since the nets 45 and 46 are formed separately from the tray 21 and the lid 31, the effect of this embodiment can be obtained by using the existing tray 21 or the lid 31. Further, the value of the height of the closed space S minus the sum of the thicknesses of the nets 45 and 46 is approximately equal to the thickness of the cell sheet-containing carrier 18 or slightly smaller than the thickness of the cell sheet-containing carrier 18. As a result, the nets 45 and 46 are brought into contact with the cell sheet-containing carrier 18 with a small contact area by the pressure that the cultured epithelial cell sheet 10 is not damaged, thereby reliably supporting the cell sheet-containing carrier 18. At the same time, it is possible to reliably prevent the cultured epithelial cell sheet 10 packaged in the cultured cell sheet package 50 from being damaged by the nets 45 and 46.

In addition, since the cultured epithelial cell sheet 10 is reinforced by the first carrier 14 formed of the wound coating material, the cultured cell sheet 10 can be effectively prevented from being damaged during handling even if the cultured cell sheet 10 does not have sufficient strength. It can be supported in an unfolded and expanded state, and the handling becomes easy, and since the cultured epithelial cell sheet 10 supported by the first carrier 14 is packaged with the second carrier 16, the cultured epithelial cell sheet ( 10) can be protected, and in particular, the base layer of the cultured epithelial cell sheet 10 having cell proliferation ability can be reliably protected. In addition, since the first carrier 14 is formed of a wound coating material, the operation of releasing the support of the first carrier 14 when the cultured epithelial cell sheet 10 is used can be easily performed.

In addition, in FIG.2 and FIG.3, the net 45 and net 46 which insert the cell sheet | seat wrapping carrier 18 are arrange | positioned so that a lattice may correspond, but you may arrange | position so that a lattice may not correspond. In addition, instead of using separate nets 45 and 46, the tray 21 and the lid 31 integrate a lattice-like wire member similar to the net 45 on the bottom surface side of the tray recess 22. Alternatively, it may be formed by molding, or a lattice-like line member similar to the net 46 may be integrally formed by molding on the rear face of the bottom face of the lead concave portion 32.

4. Preparation of Cultured Cell Sheet Package-2

4 is an assembled perspective view of the culture cell sheet package. The packaging container 60 is comprised from the tray 61, the lid 71, and the annular seal 81. As shown in FIG. The tray 61 is made of polypropylene and has a tray recess 62 for storing a liquid medium, a tray embankment 63 formed around the tray recess 62, and a tray embankment 63. The tray groove portion 64 formed along the outer circumference of the tray, the tray collar portion 65 formed along the outer circumference of the tray groove portion 64, and the bottom surface side of the tray concave portion 62 at predetermined intervals (for example, 1 cm). It is provided with the several triangular-vertical protrusion 68 provided to stand vertically and horizontally. The number of the projections 68 is determined depending on the size of the cultured epithelial cell sheet 10 and the like, the predetermined interval or the shape of the projections, so that the number of projections 68, the minimum contact area to reliably support the cultured epithelial cell sheet 10, It is preferable to determine the interval and the shape of the projections in order to suppress damage to the cultured epithelial cell sheet 10. In addition, although a vertex may be sharp, it is more preferable to be round. Moreover, the annular step 66 is formed in the inner peripheral side of the tray collar part 65, and the triangular step 67 is formed in one corner of the four corners of the tray collar part 65. As shown in FIG. On the other hand, the lid 71 is made of polypropylene and has a lid recess 72 which is shallower than the depth of the tray recess 62, a lid embankment 73 formed around the lead recess 72, and Predetermined spacing between the lead groove portion 74 formed along the outer circumference of the lead embankment portion 73, the lead collar portion 75 formed along the outer circumference of the lead groove portion 74, and the rear face rear surface of the lead recess portion 72. (For example, 1 cm), a plurality of triangular vertebral projections 78 are provided vertically and horizontally. Then, the lead recess 72 is inserted into the tray recess 62 so that the tray bank 63 and the lead bank 73 are aligned, and the tray groove 64 and the lead recess 74 are coincident with each other. When the lead collar portion 75 is superimposed on the tray collar portion 65, the lead collar portion 75 is in a state coinciding with the annular step 66 of the tray collar portion 65, and here the annular seal ( 81, the tray collar portion 65, the lead collar portion 75, and the annular seal 81 are thermally welded to form a sealed space S surrounded by the tray recess 62 and the lead recess 72. Is formed. However, when removing the annular seal 81, the annular seal 81 and the triangular step 67 are not heat-welded so that it can be removed from the triangular step 67. As shown in FIG.

In addition, the transport medium (antibiotic-containing DMEM) is placed in the tray recess 62 of the tray 61, and the cell sheet-containing carrier 18 produced in the above 2. is placed on the bottom surface side of the tray recess 62. It mounted on the some processus | protrusion 68 installed upright. Thereafter, the lead recessed portion 72 is inserted into the tray recessed portion 62, and the tray bank portion 63 and the lead bank portion 73 are aligned so that air does not remain in the sealed space S, and the tray groove portion When the 64 matches the lead groove portion 74 and the lead collar portion 75 overlaps the tray collar portion 65, the lead collar portion 75 is the annular step 66 of the tray collar portion 65. And the annular seal 81 was covered, and the tray collar portion 65, the lead collar portion 75, and the annular seal 81 were thermally welded. As a result, the sealed space S surrounded by the tray recess 62 and the lead recess 72 is filled with the transport medium, and the cell sheet-containing carrier 18 is projected into the transport medium (68, 78). It became the support state by this, and the cultured cell sheet package 90 was completed. The cross section (left half) of this culture cell sheet package 90 is shown in FIG. In addition, the above operation was performed in a clean room. In addition, the packaging container 60 used what was sterilized previously.

In this cultured cell sheet package 90, the projections 68 integrally formed on the bottom surface side of the tray recess 62 are in point contact with the cell sheet-containing carrier 18 substantially in contact with the lead recess 72. The projection 78 integrally formed on the bottom back side of the bottom surface) is in close contact with the cell sheet-containing carrier 18 to support the cell sheet-containing carrier 18 in the transport medium. In addition, the distance from the tip of the projection 68 to the tip of the projection 78 is designed to substantially coincide with the thickness of the cell sheet-containing carrier 18 or to be slightly smaller than the thickness of the cell sheet-containing carrier 18. For this reason, the force which the processus | protrusion 68,78 pressurizes the cell sheet containing carrier 18 is weak.

According to the culture cell sheet package 90 described in detail above, since the liquid medium filled in the sealed space S is almost completely removed, the culture cell sheet immersed in the liquid medium may be deteriorated by air. There is no. In addition, even if the culture cell sheet package 90 is shaken at the time of transportation, the culture medium epithelial cell sheet does not have to be shifted or missed because the bubble does not move or the bubbles do not move during the liquid medium. 18) is not easily moved or released, and there is no fear that the cultured epithelial cell sheet 10 contained in the cell sheet-containing carrier 18 collides with the tray 61 or the lid 71 and is damaged. In addition, the cell sheet-containing carrier 18 and the projections 68 and 78 are not in overall contact but merely in linear contact, and the projections 68 and 78 pressurize the cell sheet-containing carrier 18. Since the force to perform is weak, the cultured epithelial cell sheet 10 contained in the cell sheet-containing carrier 18 is hardly damaged by the projections 68 and 78. Therefore, the cultured epithelial cell sheet 10 (particularly, the base layer important for engraftment and healing of the cultured epithelial cell sheet 10) can be transported without damage.

Moreover, as shown in FIG. 5, the position where the projection 68 contacts the cell sheet-containing carrier 18 and the position where the projection 78 contacts the cell sheet-containing carrier 18 are not opposed to each other, There is no fear that excessive pressure will be applied to the cell sheet-containing carrier 18 even if deformation occurs during molding error or transportation. Moreover, since the projections 68 and 78 are integrally molded with the tray 61 and the lid 71, the operability at the time of packing or unpacking is excellent compared with the case of a separate body. Further, the value of the height of the confined space S minus the sum of the heights of the projections 68 and 78 is approximately equal to the thickness of the cell sheet-containing carrier 18 or slightly smaller than the thickness of the cell sheet-containing carrier 18. The cell sheet-containing carrier 18 is thereby reliably supported, while the projections 68 and 78 are in contact with the cell sheet-containing carrier 18 with a small contact area at a pressure at which the cultured epithelial cell sheet 10 is not damaged. It is possible to reliably prevent the cultured epithelial cell sheet 10 packaged in the cultured cell sheet package 90 from being damaged by the projections 68 and 78. The effect by the 1st carrier 14 and the 2nd carrier 16 is the same as that of the culture cell sheet package 50 of said 3.

In addition, since the tray collar portion 65 and the lead collar portion 75 are thermally welded by the annular seal 81, the culture cell sheet package 90 can be stably stacked, so that a plurality of culture cell sheets can be stacked. It can be transported easily. In this case, after placing a plate having a size that crosses the lid embankment 33 on the lid embankment 33, the other culture cell sheet package 90 is to be mounted, thereby culturing the cell sheet 10 The pressure is not applied to the enclosed space S, which is supported), so that the plurality of cultured cell sheet packages 90 can be stably stacked. In addition, you may attach such a board | plate material to the lower part of the tray recessed part 22 previously, and may integrate. Moreover, the same effect can also be acquired by making the wall surface which comprises the closed space S incline.

In addition, since the annular seal 81 is adopted, when the lid 71 is formed of a transparent member, the state in the sealed space S can be observed without opening the culture cell sheet package 90, and the Problems such as release of the carrier and damage to the culture cell sheet can be confirmed in advance. Of course, the same effect can be acquired also when both the seal 41 and the lid 31 are made into a transparent member.

In addition, instead of using the projections 68 and 78 integrally formed with the tray 61 or the lid 71, the same projection is formed on a flat plate separate from the tray 61 or the lid 71. 3. In the same manner as the nets 45 and 46, the cell sheet-containing carrier 18 may be sandwiched between these flat plates.

The present invention is not limited to the above embodiment in any way, and of course, may take various forms as long as it falls within the technical scope of the present invention. For example, in the above 3. or 4., the cultured epithelial cell sheet 10 is supported using the first carrier 14 and the second carrier 16, but the first carrier (not the second carrier 16) is used. 14) may be used, and only the second carrier 16 may be used without using the first carrier 14, and the cultured epithelial cell sheet 10 is directly netted without using both carriers 14,16. 45, 46) (or projections 68, 78). In the above 3., the tray 21 does not have the tray embankment 23 or the tray groove 24, and the tray collar 25 may be formed directly on the outer circumference of the tray recess 22, and the lead ( 31) The lead collar part 35 may be formed directly in the outer periphery of the lead recessed part 32, without having the lead bank part 33 and the lead groove part 34 (the same also about 4. said). In addition, as described in Japanese Patent Publication No. 2733800 described above, the tray collar portion 25 is formed directly on the outer circumference of the tray recess 22, and the seal 41 may be directly heat welded as a lead.

In addition, a plurality of holes may be formed in the lead groove portions 34 and 74. By doing so, a small amount of liquid medium and air removed from the sealed space S at the time of sealing pass through the gap between the tray banks 23 and 63 and the lead banks 33 and 73, and then the lead grooves 34 from the holes. And 74 are discharged above, and do not contaminate the inner circumferential side annular steps 26 and 66 of the lead collar portions 35 and 75 and the tray collar portions 25 and 65. Therefore, the seals 41 and 81 can be heat-welded easily and reliably, and the operation | work which fills a liquid medium by removing air almost completely from the sealed space S can be performed easily.

In the above-described embodiment, the tray collar portion 25, the lead collar portion 35, and the seal 41 are thermally welded, or the tray collar portion 65, the lead collar portion 75, and the annular seal ( 81) was thermally welded, but may be bonded with an adhesive or the like instead of thermal welding.

In addition, the recessed part and convex part (engaging means) as shown in FIG. 6 may be formed in four corners of the net 45 and 46, and both may be fitted, and in this case, the cell sheet | seat containing carrier 18 is a net. Since it is reliably supported between the 45 and 46, even if a slight gap is generated between the net 45 and 46 and the tray 21 or the lid 31, release of the carrier 16 and the like can be suppressed. Thus, when the recessed part and the convex part were formed in the four corners of the net 45 and 46, the cultured epithelial cell was not used without using the 2nd carrier 16 (or both of the 1st and 2nd carriers 14 and 16). By inserting the sheet 10 into the nets 45 and 46 to engage the recesses and the convex portions, the cultured epithelial cell sheet even after removing the culture epithelial cell sheet 10 together with the nets 45 and 46 from the tray 21. (10) can be reliably supported between the nets 45 and 46, and the cultured epithelial cell sheet 10 can be washed while being supported between the nets 45 and 46.

Even if it is shaken at the time of transportation by the culture cell sheet package according to the configuration of the present invention, the medium does not sway and thus there is no fear that the cultured epithelial cell sheet contained in the cell sheet-containing carrier is not displaced. It also has the effect of preventing deterioration by air.

Claims (16)

A tray made of airtight and dense material, A lid made of airtight and dense material, A liquid medium filled with air almost completely removed from the sealed space surrounded by the tray and the lid; A culture cell sheet package having a culture cell sheet immersed in the liquid medium in the sealed space. The method of claim 1, further comprising: a first interposition member between the culture cell sheet and the tray and in contact with the tray and at least partially in contact with the culture cell sheet; A culture cell sheet package comprising a second interposition member between the culture cell sheet and the lid and in contact with the lead and partially in contact with the culture cell sheet. The culture cell sheet package according to claim 2, wherein the culture cell sheet is in contact with the first interposition member and the second interposition member while being supported by a flat support. The cultured cell sheet package according to claim 3, wherein the support is formed of a sheet-like wound coating material. The method of claim 3 or 4, wherein the support is composed of a first support sheet and a second support sheet, the first support sheet is one side of the culture cell sheet is overlapping and integrated, the second support The sheet is cultured cell sheet package which is supported in a state in which the cultured cell sheet integrated with the first support sheet. The position at which the first interposition member is in contact with the culture cell sheet and the position at which the second interposition member is in contact with the culture cell sheet are not opposed to each other and are shifted apart from each other. Cultured cell sheet package. The method according to any one of claims 2 to 4, wherein the first interposition member is molded separately from the tray or integrally molded with the tray, The second interposition member is a cultured cell sheet package that is molded separately from the lead or integrally molded with the lead. The method according to any one of claims 2 to 4, wherein the first interposition member is a lattice-shaped wire member or a plurality of protrusion members, The second interposition member is a grid-shaped line member or a culture cell sheet package of the plurality of protrusion members. The method of claim 8, wherein the first interposition member is a net separate from the tray or a plurality of protrusion members integrally formed with the tray, Wherein the second interposition member is a net separate from the lead or a plurality of protrusion members integrally molded with the lead. The culture according to any one of claims 2 to 4, wherein the first interposition member and the second interposition member are brought into contact with the culture cell sheet by a pressing force such that the culture cell sheet is not damaged by both members. Cell sheet package. The cultured cell sheet package according to any one of claims 1 to 4, which is used for conveying the cultured cell sheet. The tray recess according to any one of claims 1 to 4, wherein the tray recess is formed at approximately the center of the tray; And a lead recess formed at approximately the center of the lead, The enclosed space surrounded by the tray and the lid is a space placed between the tray recess and the lead recess. The tray bank embankment of claim 12, wherein the tray embankment is formed on an outer circumference of the tray recess; A tray groove portion formed on an outer periphery of the tray embankment portion of the tray; A lead embankment portion formed on an outer periphery of the lead concave portion among the leads; A lead groove portion formed on an outer periphery of the lead embankment portion among the leads, When the sealed space is formed by the tray and the lid, the tray bank part and the lead bank part are in close contact with each other, and the tray groove part and the lead groove part are in close contact with each other. The cultured cell sheet package according to claim 13, wherein a hole is formed in at least one of the tray groove portion and the lead groove portion. The tray and the lid each have a tray collar portion and a lead collar portion, and each of the trays and the lid is attached so as to cover a seal member of a material different from that of the tray and the lid. The cultured cell sheet package thereby sealing the tray and the lid. A sheet fitting member used for a culture cell sheet package in which a culture cell sheet is immersed in a liquid medium filled with air almost completely removed from a sealed space surrounded by a tray and a lid. A first fitting member partially contacted with the culture cell sheet between the culture cell sheet and the tray and formed separately from the tray, A second fitting member partially contacted with the culture cell sheet between the culture cell sheet and the lid and formed separately from the lead; And a fitting means for engaging the first fitting member and the second fitting member substantially integrally.