JP2015119684A - Sheet-like cell culture storage container, and method of using sheet-like cell culture storage container - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sheet-like cell culture storage container from which a sheet-like cell culture can be easily taken out, and a method of using the sheet-like cell culture storage container.SOLUTION: A sheet-like cell culture storage container comprises: a culture housing container 1 which has a bottom surface 3 having a culture mounting region R for mounting a sheet-like cell culture C and a peripheral wall part 4 formed along a circumference of the bottom surface 3, in which opening parts 5 are formed in the culture mounting region R, and which is used for housing the sheet-like cell culture C together with preservation liquid S; and a communication container which communicates with the inside of the culture housing container 1 via the opening parts 5. The sheet-like cell culture C mounted on the culture mounting region R floats in the preservation liquid S by allowing the preservation liquid S to flow into the culture housing container 1 through the opening parts 5 from the communication container.

Description

  TECHNICAL FIELD The present invention relates to a sheet-shaped cell culture storage container and a method for using the sheet-shaped cell culture storage container, and in particular, a sheet-shaped cell culture storage container and a sheet-shaped cell culture containing a sheet-shaped cell culture together with a storage solution. The present invention relates to a method for using a storage container.

In recent years, regenerative medicine has been studied as a solution for treating severe heart failure such as myocardial infarction.
In severe myocardial infarction or the like, cardiomyocytes fall into dysfunction, and further, fibroblast proliferation and interstitial fibrosis progress, resulting in heart failure. As the heart failure progresses, the myocardial cells are damaged and fall into apoptosis, but the cardiomyocytes hardly undergo cell division, so the number of cardiomyocytes decreases and the cardiac function further decreases.
For such patients with severe heart failure, cell transplantation may be applied for the purpose of accelerating the recovery of the heart by sustained release of cytokines, and clinical applications using autologous skeletal myoblasts have already begun.

  In recent years, as an example of cells used for cell transplantation, by using a temperature-responsive culture dish applying tissue engineering, it has been configured in three dimensions applicable to the heart containing cells derived from parts other than the adult myocardium A cell culture and a method for producing the same have been proposed (Patent Document 1).

  However, such cell cultures are fragile because they form a relatively thin sheet. In order to detach such a sheet-shaped cell culture from the culture substrate, a mechanical technique such as pipetting and scraping, or a normal adherent cell detachment method in which enzyme treatment is performed using trypsin or the like is used. There is a possibility that the structure of the object is broken or the cells are damaged. For this reason, in Patent Document 1, cells are cultured in a culture dish coated with a temperature-responsive material, and the sheet-like cell culture is peeled off by changing the hydrophobicity of the material by changing the temperature. The method is used. However, such a culture dish is expensive, and due to the waiting time until the temperature of the culture dish decreases or various detachment states depending on the cell type, destruction of the sheet-like cell culture structure or cell detachment at the time of detachment. Problems such as damage may occur.

  Therefore, in order to gently peel the sheet-shaped cell culture, a cell culture support having a surface structure in which concave portions and convex portions are regularly arranged and whose surface is coated with a polymer has been proposed. (Patent Document 2). Cell structures produced using such cell culture supports can be gently detached from the cell culture support using hydrodynamic forces such as liquid flow or mechanical forces with instruments such as tweezers. It becomes possible to do.

Special table 2007-528755 gazette JP 2009-178050 A

  However, the sheet-shaped cell culture peeled off from the cell culture container as described above is transferred to the sheet-shaped cell culture storage container in which the storage solution is put, for example, in the storage container until immediately before the transplantation operation is performed. Because it is preserved, the sheet-shaped cell culture is sunk in close contact with the bottom of the sheet-shaped cell culture storage container when removed from the sheet-shaped cell culture storage container using a transplanter such as a spatula during transplantation. It has been difficult to insert and remove the transplant device between the bottom surface of the storage container and the sheet-like cell culture. For this reason, there is a possibility that the sheet-like cell culture may be damaged by forcibly inserting the transplant device or the removed sheet-like cell culture may be bent and hinder the transplantation operation.

  The present invention has been made to solve such a conventional problem, and uses a sheet-shaped cell culture storage container and a sheet-shaped cell culture storage container from which a sheet-shaped cell culture can be easily taken out. It aims to provide a method.

  A sheet-like cell culture storage container according to the present invention has a bottom surface having a culture placing region for placing a sheet-like cell culture, and a peripheral wall formed along the periphery of the bottom surface, and a culture. An opening is formed in the mounting region and a culture container for storing the sheet-shaped cell culture together with the preservation solution; and a communication container communicating with the inside of the culture container through the opening; The sheet-like cell culture placed in the culture placement region floats in the preservation solution by allowing the preservation solution to flow from the communication container into the culture container through the opening.

  Here, the communication container includes the culture container and has a bottom surface disposed below the bottom surface of the culture container and a peripheral wall part disposed outside the peripheral wall part of the culture container. It is preferable to have an outer container that communicates with the inside of the culture container via the opening.

  In addition, the culture container is fixed in position with respect to the outer container, and the sheet-shaped cell culture storage container is tilted and then returned to its original position, or a new storage solution is added to the outer container. It is preferable that the preservation solution in the outer container flows into the culture container through the opening by temporarily raising the water level in the outer container relative to the water level in the culture container.

  The culture container is disposed so as to be movable in the vertical direction with respect to the outer container, and the culture container is pushed down with respect to the outer container so that the water level in the outer container is temporarily higher than the water level in the culture container. By raising, the preservation solution in the outer container can also flow into the culture container through the opening.

  In addition, the communication container has a sealed chamber having a movable portion that communicates with the culture container through the opening and is filled with a storage solution and changes the internal volume. By reducing the volume, the preservation solution in the sealed chamber can flow into the culture container through the opening.

  A method for using a sheet-shaped cell culture storage container according to the present invention includes a bottom surface having a culture placing region for placing a sheet-shaped cell culture, and a peripheral wall formed along the periphery of the bottom surface. And a culture container of a sheet-like cell culture storage container, comprising: a culture container having an opening formed in the culture placement region; and a communication container communicating with the inside of the culture container through the opening. The sheet-like cell culture is placed in the culture placement region by storing the sheet-like cell culture together with the preservation solution in the container and allowing the preservation solution to flow from the communicating container into the culture accommodation container through the opening. A thing floats in a preservation solution.

  Here, the culture container is contained in the communication container and the position thereof is fixed with respect to the communication container, and the sheet-shaped cell culture storage container is used when the sheet-shaped cell culture is suspended in the preservation solution. The storage solution in the communication container can be recovered by temporarily inclining the liquid or by adding a new storage solution to the communication container and temporarily raising the water level in the communication container above the water level in the culture container. Preferably flows into the culture container through the opening.

  ADVANTAGE OF THE INVENTION According to this invention, the usage method of the sheet-like cell culture preservation container which can take out a sheet-like cell culture easily, and a sheet-like cell culture preservation container can be provided.

It is a perspective view which shows the structure of the sheet-like cell culture storage container which concerns on Embodiment 1 of this invention. The structure of the sheet-like cell culture storage container concerning Embodiment 1 is shown, (A) is a top view, (B) is sectional drawing. 3 is a cross-sectional view showing a method for using the sheet-shaped cell culture storage container according to Embodiment 1. FIG. 6 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to a modification of Embodiment 1. FIG. FIG. 6 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to another modification of the first embodiment. The structure of the sheet-like cell culture storage container concerning Embodiment 2 is shown, (A) is a top view, (B) is sectional drawing. 10 is a top view showing a configuration of a sheet-shaped cell culture storage container according to a modification of the second embodiment. FIG. The structure of the sheet-like cell culture storage container which concerns on the other modification of Embodiment 2 is shown, (A) is a top view, (B) is sectional drawing. 6 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to Embodiment 3. FIG. 6 is a cross-sectional view showing a method for using a sheet-like cell culture storage container according to Embodiment 3. FIG. 6 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to Embodiment 4. FIG. FIG. 10 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to a modification of the fourth embodiment. FIG. 10 is a cross-sectional view showing a configuration of a sheet-shaped cell culture storage container according to another modification of the fourth embodiment. It is a perspective view which shows the opening part covered with the fabric of a mesh material. It is a perspective view which shows the opening part covered with the on-off valve.

Embodiments of the present invention will be described below with reference to the accompanying drawings.
Embodiment 1
FIG. 1 shows the configuration of the sheet-shaped cell culture storage container according to Embodiment 1. This sheet-shaped cell culture storage container includes a culture container 1 for storing the sheet-shaped cell culture together with the storage solution S, and an outer container 2 disposed outside the culture container 1.

FIGS. 2A and 2B show a sheet-shaped cell culture storage container in which the sheet-shaped cell culture C is stored together with the storage solution S.
The culture container 1 has a cylindrical shape with an open top, and has a shape like a petri dish. The culture container 1 has a bottom surface 3 having a culture placement region R for placing the sheet-shaped cell culture C, and a peripheral wall portion 4 formed along the periphery of the bottom surface 3.

Here, the culture placing region R is a region in which a region on which the sheet-like cell culture C is placed is virtually set. For this reason, the culture placement region R is changed in position and range according to the size and type of the sheet-like cell culture C, and can be set near the center of the bottom surface 3, for example. it can.
A plurality of openings 5 are formed in the culture placement region R. Thereby, the some opening part 5 can be arrange | positioned so that the sheet-like cell culture C may overlap. In addition, since the position of the sheet-like cell culture C may be moved according to the shaking of the preservation solution S in the culture container 1, the culture placement region R is set over the entire bottom surface 3. Even if the plurality of openings 5 are formed over the entire bottom surface 3 and the position of the sheet-shaped cell culture C is moved within the culture container 1, any one of the openings 5 is located immediately below the sheet-shaped cell culture C. It is preferable to arrange so that it exists. Moreover, it is preferable that the plurality of openings 5 are each formed with a diameter that does not allow the sheet-like cell culture C to enter the opening 5 greatly, for example, with a diameter of 0.1 mm to 2.0 mm. .

The outer container 2 has a petri dish-like shape like the culture container 1, and contains the culture container 1 in the center thereof. In addition, the outer container 2 has a bottom surface 6 disposed below the bottom surface 3 of the culture container 1 and a peripheral wall part 7 disposed outside the peripheral wall part 4 of the culture container 1. The position of the culture container 1 is fixed with respect to the outer container 2 by a fixing portion 8 that connects the culture container 1 and the outer container 2. As described above, the culture container 1 is disposed at a distance from the outer container 2 so that the outer container 2 communicates with the inside of the culture container 1 through the plurality of openings 5. In addition, a liquid reservoir 9 for storing the preservation solution S is formed outside the culture container 1. That is, the liquid reservoir 9 is formed so as to cover the peripheral wall 4 and the bottom surface 3 of the culture container 1 from the outside, and an annular receiving port 10 between the upper part of the peripheral wall 4 and the upper part of the peripheral wall 7. Have
Here, the outer container 2 communicates with the inside of the culture container 1 through a plurality of openings 5, and constitutes the communication container of the present invention.

Thus, the preservation solution S is stored in the culture container 1 and the outer container 2 that communicate with each other through the plurality of openings 5. Further, a sheet-shaped cell culture C is stored in the culture storage container 1 in which the preservation solution S is stored.
The sheet-shaped cell culture C accommodated in the culture container 1 is not particularly limited as long as it is a cell cultured in a sheet shape. For example, myoblasts (for example, skeletal muscles) Blast cells), cardiomyocytes, fibroblasts, synoviocytes, epithelial cells, endothelial cells and the like.

Next, the usage method of a sheet-like cell culture preservation container is demonstrated.
First, the sheet-shaped cell culture C cultured in a sheet shape in a cell culture container or the like is peeled off from the cell culture container, and then transferred to the culture container 1 of the sheet-shaped cell culture storage container. It is preserve | saved with the preservation | save liquid S in the culture container 1 until just before a transplant operation | work. The sheet-like cell culture C transferred into the culture container 1 sinks in the preservation solution S and is set on the bottom surface 3 of the culture container 1 as shown in FIGS. 2 (A) and (B). Is placed on the culture placement region R.

  In general, the sheet-shaped cell culture C is taken out of the culture container 1 using an implantation device such as a spatula. However, as described above, the sheet-shaped cell culture C sinks in the preservation solution S and is bottom. When placed on 3, it becomes difficult to insert and remove the transplant device between the sheet-shaped cell culture C and the bottom surface 3.

Then, when taking out the sheet-shaped cell culture C from the culture container 1, the operator once tilts the sheet-shaped cell culture storage container as shown in FIG. Thereby, the preservation solution S in the culture container 1 is transferred from the upper part of the culture container 1 into the liquid reservoir 9 through the receiving port 10 of the outer container 2.
Here, the peripheral wall portion 7 of the outer container 2 preferably extends higher than the peripheral wall portion 4 of the culture container 1. Thereby, since the preservation | save liquid S moved from the culture container 1 into the outer container 2 is reliably received by the surrounding wall part 7 of the outer container 2, it transfers to the outer container 2 reliably, without reducing the preservation | save liquid S. be able to. In addition, since the culture container 1 is contained in the center of the outer container 2, the liquid reservoir 9 of the outer container 2 is formed so as to surround the culture container 1, and preserves the sheet-shaped cell culture. Even if the container is inclined in any direction, the preservation solution S in the culture container 1 can be transferred into the outer container 2.

Subsequently, when the operator returns the inclined sheet-shaped cell culture storage container to the original state, the water level L1 in the outer container 2 is changed to the water level L2 in the culture container 1 as shown in FIG. Is raised more than. For this reason, as shown in FIG. 3C, the preservation solution S in the outer container 2 is cultured through the plurality of openings 5 until the water level L1 in the outer container 2 and the water level L2 in the culture container 1 match. It gently flows into the object storage container 1.
Here, the plurality of openings 5 are formed in the culture placement region R on which the sheet-like cell culture C is placed. For this reason, the preservation solution S gently flows into the culture container 1 from the outer container 2 through the plurality of openings 5, whereby the preservation solution S is placed on the culture placement region R. The back side of the cell culture C can be gradually pushed up, and the sheet-like cell culture C can be suspended in the preservation solution S.

As a result, a gap G for inserting the transplant device D is formed between the sheet-like cell culture C suspended in the preservation solution S and the bottom surface 3 of the culture container 1, so that FIG. As shown in FIG. 2, the transplant device D can be smoothly inserted between the sheet-shaped cell culture C and the bottom surface 3 of the culture container 1.
The plurality of openings 5 are preferably arranged so as to overlap the entire surface of the sheet-like cell culture C, but may be arranged so as to overlap a part of the sheet-like cell culture C. Even when the plurality of openings 5 overlap with a part of the sheet-like cell culture C, at least a part of the sheet-like cell culture C can be suspended. Even when only a part of the sheet-like cell culture C floats, the transplant device D can be inserted between the sheet-like cell culture C and the bottom surface 3 starting from the part.

  In this way, the sheet-like cell culture C placed on the transplant device D is taken out from the culture container 1 and used for cell transplantation.

  According to this embodiment, in order to gently flow the preservation solution S into the culture container 1 due to the difference in water level generated between the culture container 1 and the outer container 2, the sheet-shaped cell culture C is It can be suspended in the preservation solution S without being damaged. In addition, the sheet-shaped cell culture C is suspended in the preservation solution S to suppress the bending of the sheet-shaped cell culture C, and the sheet-shaped cell culture C and the culture container 1 are expanded in a state where the whole is expanded. A gap G can be formed between the bottom surface 3. Furthermore, the sheet-like cell culture C can be easily suspended in the preservation solution S simply by tilting the sheet-like cell culture storage container, and the sheet-like cell culture C and the bottom 3 of the culture container 1 The transplant device D can be smoothly inserted into the gap G. Thereby, since the sheet-shaped cell culture C is taken out in a state in which the damage of the sheet-shaped cell culture C is suppressed and the whole is expanded, the cell-transplanting operation can be smoothly proceeded.

  According to the above embodiment, the water level L1 in the outer container 2 is temporarily raised above the water level L2 in the culture container 1 by tilting the sheet-shaped cell culture storage container and then returning it to the original state. However, it is sufficient that a water level difference is generated between the outer container 2 and the culture container 1, and the present invention is not limited to this.

For example, the sheet-shaped cell culture storage container shown in FIGS. 2A and 2B is newly stored in the liquid reservoir 9 of the outer container 2 using a pipette P or the like as shown in FIG. By adding the liquid S, the water level L1 in the outer container 2 can be temporarily raised above the water level L2 in the culture container 1. Thereby, the preservation solution S in the outer container 2 gently flows into the culture container 1 through the plurality of openings 5, and the sheet-like cell culture C in the culture container 1 floats in the preservation solution S. Can be made.
Thus, when the sheet-shaped cell culture C is suspended in the preservation solution S, it is not necessary to incline the sheet-shaped cell culture storage container. It can be stably held in R. For this reason, sheet-like cell culture C does not move and wrinkles or the like occur, or sheet-like cell culture C does not move from culture container 1 to outer container 2 together with preservation solution S. The cell culture C can be smoothly taken out from the culture container 1.

Further, as shown in FIG. 5, the outer container 2 has a peripheral wall portion 13 formed so that the lower portion 11 is close to the peripheral wall portion 4 of the culture preservation container 1 and the upper portion 12 is greatly opened from the lower portion 11. be able to. For this reason, the receiving port 10 of the liquid reservoir 9 is formed wide between the peripheral wall 4 and the upper portion 12 of the peripheral wall 13, and a part of the liquid reservoir 9 extending downward from the receiving port 10 is part of the peripheral wall 4. And a lower portion 11 of the peripheral wall portion 13.
As a result, when the sheet-shaped cell culture storage container is tilted, the storage liquid S in the culture storage container 1 is reliably received in the receiving port 10 formed wide, so the storage liquid S is reduced. It can be reliably transferred to the outer container 2 without any problem. Further, when the inclined sheet-shaped cell culture storage container is returned to its original state, the storage liquid S received in the receiving port 10 gradually passes through the narrowly formed portion of the liquid reservoir 9. Therefore, the preservation solution S can be allowed to flow into the culture preservation container 1 over time, whereby the sheet-like cell culture C can be suspended for a long time. For this reason, the transplant device D can be steadily inserted between the sheet-shaped cell culture C and the bottom surface 3 of the culture container 1.

Embodiment 2
In the first embodiment, the culture container 1 is disposed so as to be included in the central portion of the outer container 2, but the culture container 1 only needs to be included in the outer container and is not limited thereto. Absent.

  For example, as shown in FIGS. 6 (A) and 6 (B), an outer container 21 that encloses the culture container 1 so as to be offset to one side can be provided. The outer container 21 includes a bottom surface 22 disposed below the bottom surface 3 of the culture container 1 and a wall 23 disposed outside the half surface of the peripheral wall 4 of the culture container 1. Have. That is, the bottom surface 22 is disposed so as to cover the entire bottom surface 3 of the culture container 1, and the wall portion 23 is disposed so as to cover the half surface of the peripheral wall portion 4 of the culture container 1. Further, the outer container 21 communicates with the inside of the culture container 1 through the plurality of openings 5, and the preservation solution S is stored in the inside. As a result, a liquid reservoir 24 for storing the preservation solution S is formed between the outer container 21 and the culture container 1. The liquid reservoir 24 is formed so as to cover the entire bottom surface 3 of the culture container 1 and the half surface of the peripheral wall 4, and a receiving port between the upper part of the peripheral wall 4 and the upper part of the wall 23. 25.

Next, a method of using the sheet-like cell culture storage container shown in FIGS. 6 (A) and 6 (B) will be described.
When taking out the sheet-shaped cell culture C from the sheet-shaped cell culture storage container, the sheet-shaped cell culture storage container has the wall 23 side of the outer container 21 facing downward and the opposite side of the wall 23 facing upward. The storage solution S in the culture container 1 is once inclined so as to be transferred into the liquid reservoir 24 of the outer container 21 through the receiving port 25 and then returned to the original state. As a result, the water level in the outer container 21 is raised above the water level in the culture container 1, so that the preservation solution S in the outer container 21 flows into the culture container 1 through the plurality of openings 5.

  At this time, the liquid reservoir 24 is formed so as to cover only the half surface of the peripheral wall 4 of the culture storage container 1, and the storage liquid S transferred from the culture container 1 is stored in the back of the liquid reservoir 24. It is blocked by the portion 24 a and all of it is directed to the plurality of openings 5. For this reason, the preservation | save liquid S moved in the outer side container 21 can be made to flow in into the culture | cultivation accommodation container 1 through the several opening part 5 without loss, and the sheet-like cell culture C is reliably contained in the preservation | save liquid S. Can be floated.

  According to the present embodiment, since the liquid reservoir 24 is formed so as to cover only the half surface of the peripheral wall 4 of the culture storage container 1, after the sheet-shaped cell culture storage container is once inclined, When returned, the preservation solution S transferred into the outer container 21 can be directed to the plurality of openings 5 without loss, and the sheet-shaped cell culture C stored in the culture container 1 can be reliably obtained. Can be suspended.

  A spout for pouring the preservation solution S into the receiving port 25 of the outer container 21 is provided at the edge of the peripheral wall 4 of the culture container 1 at a position facing the wall 23 of the outer container 21. It is preferable. For example, as shown in FIG. 7, the spout 26 which has a triangular cross section and a side part converges toward a front-end | tip part can be provided. Thereby, when the sheet-shaped cell culture storage container is tilted, the storage liquid S in the culture container 1 can be easily transferred into the outer container 21. Moreover, since the preservation | save liquid S in the culture container 1 can be poured in the outer container 21 in the narrow range from the front-end | tip part of the spout 26, the receiving port 25 of the outer container 21 can be formed small. Thus, by forming the receiving port 25 small, the preservation solution S transferred into the outer container 21 can be further directed to the plurality of openings 5 without loss, and is stored in the culture container 1. The sheet-shaped cell culture C can be suspended more reliably.

The outer container 21 is connected to the receiving port 25 of the outer container 21 and is provided with a liquid receiving portion extending along the outer periphery of the peripheral wall portion 4 of the culture container 1, so When the preservation solution S is transferred to the other portion, it is preferable to guide the preservation solution S to the receiving port 25 by the solution receiving portion.
For example, as shown in FIGS. 8A and 8B, the liquid receiving portion 27 can be provided so as to surround the peripheral wall portion 4 of the culture container 1 together with the receiving port 25. The liquid receiving portion 27 includes a bottom surface 28 connected to the receiving port 25 from both sides, and a wall portion connected to both side portions of the wall portion 23 and disposed so as to surround the outer peripheral wall portion 4 of the culture container 1. 29. Since the liquid receiver 27 guides the storage solution S to the receiving port 25, the storage solution S is transferred from the culture storage container 1 to the outer container 21 regardless of the direction of inclination of the sheet-shaped cell culture storage container. be able to. For this reason, the operator does not need to pay attention to the direction in which the sheet-shaped cell culture storage container is tilted when transferring the storage solution S from the culture storage container 1 to the outer container 21, thereby stabilizing the work. Can proceed.

Embodiment 3
In Embodiments 1 and 2, the position of the culture storage container is fixed with respect to the outer container. However, the culture storage container may be arranged so as to be movable in the vertical direction with respect to the outer container.

For example, as shown in FIG. 9, an elastic member 31 is provided that elastically connects the culture storage container 1 and the outer container 2 in place of the fixing portion 8 in the sheet-shaped cell culture storage container of the first embodiment. The culture preservation container 1 can be arranged so as to be movable in the vertical direction with respect to the outer container 2.
When taking out the sheet-shaped cell culture C from the sheet-shaped cell culture storage container, as shown in FIG. 10, the culture storage container 1 is pushed down with respect to the outer container 2, and the water level L1 in the outer container 2 is set to the culture. It is temporarily raised above the water level L2 in the storage container 1. As a result, the storage solution S in the outer container 2 gently flows into the culture storage container 1 through the plurality of openings 5 until the water level L1 in the outer container 2 and the water level L2 in the culture container 1 match. The sheet-shaped cell culture C placed in the culture placement region R on the bottom surface 3 of the culture preservation container 1 is suspended in the preservation solution S.

  According to the present embodiment, when the sheet-shaped cell culture C is suspended in the preservation solution S, it is not necessary to incline the sheet-shaped cell culture storage container, so that the position of the sheet-shaped cell culture C is cultured. The object can be stably held in the object placement region R. For this reason, sheet-like cell culture C does not move and wrinkles or the like occur, or sheet-like cell culture C does not move from culture container 1 to outer container 2 together with preservation solution S. The culture C can be smoothly taken out from the culture container 1.

Embodiment 4
In the first to third embodiments, the communication container is composed of an outer container that encloses the culture container 1, but the communication container only needs to communicate with the inside of the culture container 1 through the opening. However, it is not limited to this. For example, the communication container has a sealed chamber having a movable part that communicates with the culture container through a plurality of openings and is filled with a storage solution and changes the internal volume. By reducing the internal volume, the preservation solution in the sealed chamber can be allowed to flow into the culture container through the opening.

For example, as shown in FIG. 11 (A), a communication container 41 is arranged below the bottom surface 3 of the culture storage container 1 in place of the outer container 2 in the sheet-shaped cell culture storage container of the first embodiment. Can do.
The communication container 41 has a bottom surface 42 disposed below the bottom surface 3 of the culture storage container 1 and a peripheral wall portion 43 disposed on an extension line from the lower end of the peripheral wall portion 4 of the culture storage container 1. The bottom surface 42 and the peripheral wall portion 43 are connected to each other. As a result, the communication container 41 is formed with a sealed chamber 44 surrounded by the bottom surface 3, the bottom surface 42, and the peripheral wall portion 43, and the sealed chamber 44 communicates with the culture storage container 1 through the plurality of openings 5. The interior is filled with the storage solution S.
Further, the bottom surface 42 of the communication container 41 includes a movable part 45 that can be expanded and contracted, and the internal volume of the sealed chamber 44 can be changed by expanding and contracting the movable part 45. The movable portion 45 may be provided on a part of the bottom surface 42 without being formed on the entire bottom surface 42 of the communication container 41.

  When taking out the sheet-shaped cell culture C from the sheet-shaped cell culture storage container, as shown in FIG. 11B, the operator presses the movable portion 45 from the outside to reduce the internal volume of the sealed chamber 44. Let As a result, the storage solution S in the sealed chamber 44 is pushed into the culture storage container 1 through the plurality of openings 5, and the storage solution S in the sealed chamber 44 can flow into the culture storage container 1. Then, the sheet-like cell culture C placed in the culture placement region R on the bottom surface 3 of the culture preservation container 1 floats in the preservation liquid S by the preservation solution S flowing into the culture preservation container 1. Is done.

  According to the present embodiment, since the sealed chamber 44 is reduced according to the pressing of the movable portion 45 by the operator, the water pressure of the storage solution S flowing into the culture storage container 1 from the plurality of openings 5 can be easily set. It is possible to control the amount of floating of the sheet-shaped cell culture C easily.

In the third embodiment, the movable portion 45 is disposed on the bottom surface 42 of the communication container 41. However, the movable portion 45 is not limited to this as long as the internal volume of the sealed chamber 44 can be changed.
For example, as shown in FIG. 12, the movable portion 46 can be disposed on the peripheral wall portion 47 of the communication container 41. That is, the communication container 41 includes a rigid bottom surface 48 provided in place of the elastically deformable bottom surface 45, and a peripheral wall portion 47 connected to the bottom surface 48 and having rigidity. The movable part 46 is disposed in the part. When taking out the sheet-shaped cell culture storage container C from the sheet-shaped cell culture storage container, the operator presses the movable part 46 from the side with the sheet-shaped cell culture storage container placed on the support base. Thus, the internal volume of the sealed chamber 44 is reduced. As a result, the storage solution S in the sealed chamber 44 is caused to flow into the culture storage container 1 through the plurality of openings 5, and the sheet-shaped cell culture placed in the culture placement region R of the culture storage container 1. The substance C floats in the preservation solution S.
Thus, since the movable part 46 can be pressed in a state where the sheet-shaped cell culture storage container is placed on the support base, the position of the sheet-shaped cell culture C is stably within the culture placing region R. Can be fastened. For this reason, it can suppress that the sheet-like cell culture C moves and wrinkles generate | occur | produce, and can take out the sheet-like cell culture C from the culture container 1 smoothly.

In addition, the movable portion can be disposed outside the peripheral wall portion of the communication container 41.
For example, as shown in FIG. 13, the communication container 41 includes a rigid bottom surface 49 and a peripheral wall portion 50, and a movable portion 51 including a syringe detachably connected to the outside of the peripheral wall portion 50. can do. The movable portion 51 accommodates the storage solution S therein and communicates with the inside of the peripheral wall portion 50 of the communication container 41. That is, the inside of the movable portion 51 constitutes a part of the sealed chamber 44.
When taking out the sheet-shaped cell culture storage C from the sheet-shaped cell culture storage container, the operator moves the movable part by pressing the plunger of the syringe while the sheet-shaped cell culture storage container is placed on the support base. 51 is pressed to reduce the internal volume of the movable portion 51, that is, a part of the sealed chamber 44. As a result, the preservation solution S in the movable portion 51 is supplied into the sealed chamber 44 disposed inside the peripheral wall portion 50, and the preserved solution S in the pushed-out sealed chamber 44 is stored in the culture through the plurality of openings 5. The sheet-like cell culture C that flows into the container 1 and is placed in the culture placement region R of the culture preservation container 1 floats in the preservation solution S.

  Thus, since the movable part 51 can be pressed in a state where the sheet-shaped cell culture storage container is placed on the support base, the position of the sheet-shaped cell culture C is stably within the culture placement region R. It is possible to suppress the occurrence of wrinkles due to movement of the sheet-shaped cell culture C. Further, by disposing the movable portion 51 outside the peripheral wall portion 50 of the communication container 41, the height of the communication container 41 can be lowered, and accordingly, the height of the sheet-like cell culture container is lowered. be able to. For this reason, hindering cell transplantation, for example, preventing the operator's arm from coming into contact with the upper part of the sheet-like cell culture storage container can be carried out, and the cell transplantation can proceed smoothly. Furthermore, by connecting the movable part 51 to the communication container 41 in a detachable manner, the movable part 51 may be attached only when the sheet-shaped cell culture C is taken out, and the sheet-shaped cell culture storage container can be made compact. it can.

  In addition, the movable part 51 should just be arrange | positioned on the outer side of the surrounding wall part 50, can change the internal volume, and can supply the preservation | save liquid S in the sealed chamber 44, and is not restricted to a syringe. For example, a bag-like movable part containing the storage liquid S can be provided inside, and the internal volume is reduced by pressing the movable part so as to be crushed from the outside, so that the storage liquid S is placed in the sealed chamber 44. Can be injected.

Moreover, in Embodiment 1-4, the opening part 5 can be made into a structure where the sheet-like cell culture C does not enter into the opening part 5 through the preservation | save liquid S which flows in from a communicating container. .
For example, as shown in FIG. 14, the opening 5 is covered with a mesh material dough 61 so that the preservation solution S flowing from the communication container is passed and the sheet-shaped cell culture C does not enter the opening 5. can do. Moreover, as shown in FIG. 15, the opening part 5 can also be covered with the on-off valve 62 provided so that opening and closing was possible. The on-off valve 62 is opened by the hydraulic pressure of the storage solution S flowing from the communication container, and thus allows the storage solution S flowing from the communication container to pass therethrough so that the sheet-shaped cell culture C does not enter the opening 5. Can be.
In this way, the sheet-shaped cell culture C placed on the bottom of the culture storage container is prevented from entering the opening 5, whereby damage to the sheet-shaped cell culture C can be suppressed.

Moreover, in Embodiment 1-4, although the opening part 5 is formed in circular shape, what is necessary is just to be able to connect the inside of a communicating container and the inside of a culture storage container, It is not restricted to this. For example, it can be formed in a rectangular shape or a line shape.
Moreover, in Embodiment 1-4, although the several opening part 5 was formed in the culture | cultivation mounting area | region R of the culture container, the sheet-like cell culture by the preservation | save liquid S which flows in from a communicating container. It is sufficient if C can be suspended in the preservation solution S, and one opening can be formed in the culture placement region R.

  DESCRIPTION OF SYMBOLS 1 Culture container, 2,21 Outer container, 3 Bottom face, 4 Perimeter wall part, 5 Opening part, 6, 22, 42, 48, 49 Bottom face, 7, 13, 43, 47, 50 Perimeter wall part, 8 Fixing part, 9,24 Liquid reservoir, 10,25 Receiving port, 11 Lower part of peripheral wall part, 12 Upper part of peripheral wall part, 23 Wall part, 26 Spout, 27 Liquid receiving part, 28 Bottom surface, 29 Wall part, 31 Elastic member, 41 Communication container, 44 closed chamber, 45, 46, 51 movable part, 61 dough, 62 open / close valve, C sheet-like cell culture, S preservation solution, R culture mounting area, L1 water level in outer container, L2 culture accommodation Water level in the container, D transplant device, G gap, P pipette.

Claims (7)

A sheet having a bottom surface having a culture placement region for placing a sheet-shaped cell culture and a peripheral wall formed along a periphery of the bottom surface, and having an opening formed in the culture placement region and a sheet A culture container for containing a cell culture together with a preservation solution;
A communication container communicating with the inside of the culture container via the opening;
With
A sheet in which the sheet-like cell culture placed in the culture placement region floats in the preservation solution by allowing the preservation solution to flow from the communication container into the culture container through the opening. Cell culture storage container.   The communication container includes the culture container and has a bottom surface disposed below the bottom surface of the culture container and a peripheral wall part disposed outside the peripheral wall part of the culture container. The sheet-shaped cell culture storage container according to claim 1, further comprising an outer container that communicates with the inside of the culture container through the opening.   The culture container is fixed in position with respect to the outer container, and the sheet-shaped cell culture storage container is tilted and then returned to the original one, or the storage liquid is newly added to the outer container. The storage solution in the outer container flows into the culture container through the opening by adding and temporarily raising the water level in the outer container to be higher than the water level in the culture container. The sheet-like cell culture storage container according to 2. The culture container is arranged to be movable in the vertical direction with respect to the outer container,
By pushing down the culture container with respect to the outer container to temporarily raise the water level in the outer container relative to the water level in the culture container, the preservation solution in the outer container passes through the opening. The sheet-like cell culture preservation container according to claim 2, which flows into the culture container. The communication container has a sealed chamber having a movable part that communicates with the culture container through the opening and is filled with the preservation solution and changes the internal volume.
The sheet-like cell culture storage container according to claim 1, wherein the storage liquid in the sealed chamber flows into the culture container through the opening by reducing the internal volume of the sealed chamber by the movable part. . A culture having a bottom surface having a culture placement region for placing a sheet-shaped cell culture and a peripheral wall formed along the periphery of the bottom surface, and having an opening formed in the culture placement region A sheet-like cell culture together with a preservation solution in a culture-like container of a sheet-like cell culture preservation container comprising: a substance-contained container; and a communication container that communicates with the inside of the culture-contained container via the opening. Contain,
The sheet-like cell culture placed in the culture placing region floats in the preservation solution by allowing the preservation solution to flow from the communication container into the culture container through the opening. To use a sheet-like cell culture storage container.   The culture container is contained in the communication container, and the position of the culture container is fixed with respect to the communication container. When the sheet cell culture is suspended in the storage solution, the sheet cell culture By tilting the material storage container and then returning it to the original position, or by newly adding the storage solution to the communication container and temporarily raising the water level in the communication container relative to the water level in the culture container The method for using a sheet-shaped cell culture storage container according to claim 6, wherein the storage solution in the communication container flows into the culture container through the opening.

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