JP2012044872A - Cell culture vessel, cell culture apparatus, cell culture system and cell culture method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent disturbance of observation of cultured cells caused by occurrence of wrinkles in a cell culture vessel in a cell culture using the cell culture vessel composed of a soft packing material.SOLUTION: The cell culture vessel in which cells and a culture solution are sealed and which is composed of a soft packing material for performing cell culture includes a means for preventing occurrence of wrinkles. The means for preventing occurrence of wrinkles is a frame or a plate having rigidity or a part formed in thickness to have rigidity in the cell culture vessel. A loading table for loading the cell culture vessel is equipped with a means for preventing occurrence of wrinkles of the cell culture vessel and the cell culture vessel may be equipped with an area for preventing occurrence of wrinkles by the means for preventing occurrence of wrinkles. Further, the circumference of a region for observing cells in the cell culture vessel and the circumference of an observation hole in the loading table are each equipped with connection means that are mutually connected and fix the cell culture vessel on the loading table and the area for preventing occurrence of wrinkles is provided with the region for observing cells by connecting the cell culture vessel to the loading table.

Description

  The present invention relates to a cell culture technique, and more particularly to a cell culture container, a cell culture apparatus, a cell culture system, and a cell culture method capable of appropriately observing cultured cells in cell culture using a cell culture container made of soft packaging material. About.

In recent years, in the fields of pharmaceutical production, gene therapy, regenerative medicine, immunotherapy, and the like, it has been required to efficiently culture a large amount of cells, tissues, microorganisms, and the like in an artificial environment.
Under such circumstances, a large amount of cells are automatically cultured using a cell culture container made of soft packaging material. For example, Patent Document 1 discloses an automatic device comprising a culture vessel formed of a soft wrapping material and a culture device that can automatically control the culture volume according to the number of cells by monitoring the number of cells in the culture vessel with a CCD camera. A culture system is described. Thus, when automatically mass-culturing cells, a mechanism for monitoring the number and state of the cultured cells and adjusting the culture conditions to an optimum environment is indispensable. A mechanism for automatically counting cells based on an image captured by a CCD camera is one of the most effective monitoring methods.

  In addition, in order to improve the cell culture efficiency, the culture system is also required to have a mechanism for stirring the cells and the culture solution homogeneously. In the culture system of Patent Document 1, a mechanism for stirring the inside of the culture container by rotating a roller that contacts the culture container made of soft packaging material, and in Patent Document 2, a protrusion is repeatedly pressed against a flexible culture bag. Thus, a mechanism for shaking and stirring the inside of the container is described.

International Publication No. 2008/136371 Pamphlet International Publication No. 2007/052718 Pamphlet Special table 2004-514432 gazette

  However, when the cell culture vessel is made of a soft wrapping material, if the culture solution is stirred, the surface may be wrinkled or a refraction mark may be generated. As a result, wrinkles appear in the photograph obtained by automatically photographing the inside of the cell culture container, and the number of cells cannot be counted accurately, and the state of the cells cannot be observed accurately.

  Therefore, as a result of intensive studies, the present inventors have provided a means for preventing the generation of wrinkles such as a rigid frame or plate in a cell culture container made of soft packaging material, or a loading table on which the cell culture container is placed. The above-described problem is solved by providing a cell culture device equipped with a means for preventing the generation of wrinkles in the cell culture container, or by providing a means for preventing the occurrence of wrinkles in both the cell culture container and the cell culture device. Successfully completed the present invention.

Here, in the cell culture container and the cell culture device, those provided with means for preventing the occurrence of wrinkles are not found in the prior art documents, but as a technical document describing the cell culture device provided with the frame, a patent document 3 can be mentioned.
However, this cell culture vessel is entirely constituted by a frame, is not made of a soft wrapping material, and has rigidity, and prevents generation of wrinkles on the surface of the flexible cell culture vessel as in the present invention. It was not applicable.
The present invention has been made in view of such circumstances, and wrinkles are generated in an observation region for observing cells in a cell culture container by providing the cell culture container with a means for preventing the generation of wrinkles. It is an object of the present invention to provide a cell culture container, a cell culture device, a cell culture system, and a cell culture method that can prevent this.

  In order to achieve the above object, the cell culture container of the present invention is a cell culture container made of a soft wrapping material, in which cells and a culture solution are enclosed, and for culturing the cell. A means for preventing the occurrence of wrinkles is provided, and a region in which the generation of wrinkles is prevented is formed in at least a part of the cell culture container by the means for preventing the occurrence of wrinkles.

  The cell culture device of the present invention is a cell culture device provided with a loading table on which a cell culture vessel made of soft packaging material is placed for enclosing cells and a culture solution and culturing the cells. The loading table is provided with means for preventing the generation of wrinkles in the cell culture container, and the cell culture container is placed on the loading table and the generation of wrinkles in at least a part of the cell culture container by means of preventing the generation of wrinkles. In this configuration, a region in which the above is prevented is formed.

  The cell culture system of the present invention includes a cell culture container made of soft packaging material for enclosing cells and a culture solution and culturing the cell, and a cell culture having a loading table on which the cell culture container is placed. The system includes coupling means for coupling the cell culture container to the loading table by bonding to each other around the area for observing cells in the cell culture vessel and around the observation hole in the loading table. By combining, the region for observing cells is formed in a region where wrinkles are prevented.

  The cell culture method of the present invention is a cell culture method for culturing cells by enclosing cells and a culture solution in a cell culture container made of a soft wrapping material, and means for preventing the generation of wrinkles in the cell culture container A region in which the generation of wrinkles is prevented by at least a part of the cell culture container by means for preventing the generation of wrinkles, and the cells in the cell culture container are observed through this region .

  The cell culture method of the present invention is a cell culture method in which cells and culture solution are sealed in a cell culture container made of soft packaging material, and the cells are cultured. The cell culture container is placed on the loading platform, and an area in which the generation of wrinkles is prevented is formed in at least a part of the cell culture container by the means for preventing the generation of wrinkles. This is a method for observing cells in a cell culture vessel.

  The cell culture method of the present invention is a cell culture method for culturing cells by enclosing cells and a culture solution in a cell culture container made of a soft wrapping material, and is an area for observing cells in the cell culture container Are connected to each other around the observation hole and the observation hole in the loading table to fix the cell culture container to the loading table. This is a method of forming a region where generation is prevented and observing cells in the cell culture container through this region.

  ADVANTAGE OF THE INVENTION According to this invention, it can prevent that a wrinkle generate | occur | produces in the area | region for observing the cell in a cell culture container. For this reason, when culturing cells, it is possible to appropriately observe the number and state of cells in the cell culture container.

It is a figure which shows schematic structure of the cell culture container of this invention. It is a figure which shows schematic structure of the cell culture system of this invention. It is a figure which shows the longitudinal cross-section of schematic structure of the cell culture system of this invention. It is a figure which shows the structure of the cell culture container in 1st embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the external appearance and partial expansion of the cell culture container in 1st embodiment of this invention. It is a figure which shows the structure of the cell culture container in 2nd embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture container in 3rd embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture apparatus in 4th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture apparatus in 5th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture apparatus in 6th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture apparatus in 7th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture apparatus in 8th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture system in 9th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows the structure of the cell culture system in 10th embodiment of this invention, (a) is the front view, (b) is the top view. It is a figure which shows schematic structure of the conventional cell culture container. It is a figure which shows the longitudinal cross-section of schematic structure of the conventional cell culture system.

Hereinafter, embodiments of the present invention will be specifically described.
In general, in cell culture using a cell culture container, in order to observe the number and state of the cultured cells, the inside of the cell culture container is photographed through the surface of the cell culture container.
In the present invention, wrinkles are prevented from occurring in an area for observing cultured cells in such a cell culture vessel (hereinafter referred to as an observation area), and wrinkles are prevented from appearing in a photograph taken. This makes it possible to properly observe the cells.

As means for preventing the occurrence of such wrinkles, as will be described in the following embodiments, a cell culture container, a cell culture apparatus equipped with a loading table, and a cell culture container and a stack Mentioned for both of the stands.
First, the schematic configuration of the present invention will be described with reference to FIG. 1 to FIG. 3 by taking as an example one provided with a cell culture container for preventing generation of wrinkles. 1 to 3 are diagrams showing a schematic configuration of the cell culture container of the present invention, a schematic configuration of a cell culture system having the cell culture container, and a longitudinal section of the schematic configuration of the system.

As shown in FIG. 1, the cell culture container 10 of the present embodiment may include a fixing member 11. In the example of the figure, a rigid frame made of, for example, a resin is attached to a part of one side surface of the cell culture container 10. By sticking such a frame on the surface of the cell culture container 10, tension is applied to the portion surrounded by the frame on the surface of the cell culture container 10 so that wrinkles are not generated.
Thus, even when the ridge 12 is generated on the surface of the cell culture container 10 by, for example, stirring the culture solution in the cell culture container 10, the ridge 12 is applied to the observation region stretched by the tension applied by the fixing member 11. Never get in.

  FIG. 2 shows a cell culture system having a cell culture apparatus in which such a cell culture container 10 is placed on a loading table 20. The cell culture container 10 is placed on the loading table 20 and fixed by the container fixture 22 so that the observation region is arranged on the observation hole 21 in the loading table 20. The illumination device 40 is disposed above the observation region and irradiates the observation region with light, and the microscope 30 is configured to photograph the cells in the cell culture container 10 through the observation hole 21.

FIG. 3 is an enlarged cross-sectional view of a part of such a cell culture system as viewed from the side. The observation region in the cell culture container 10 is provided with a fixing member 11, thereby forming a region where wrinkles are prevented from occurring on the lower surface of the cell culture container 10. The cell culture vessel 10 is filled with a culture solution 13 and cells 14. The microscope 30 can photograph the cells 14 in the cell culture container 10 through the observation hole 21 in the loading table 20.
Thus, the cells in the cell culture container 10 can be automatically photographed through the observation region where the generation of wrinkles is prevented by the fixing member 11, and the number and situation of the cells can be appropriately observed. .

On the other hand, FIG. 15 shows a state of a conventional cell culture container 100. When the cocoon 120 is generated in such a cell culture container 100, the cocoon 120 is generated in the observation region as shown in FIG. FIG. 16 is an enlarged cross-sectional view of a part of a cell culture system having a cell culture apparatus in which such a conventional cell culture container 100 is placed on a loading table 200 as viewed from the side. The cell culture container 100 is filled with a culture solution 130 and cells 140. When the cell culture vessel 100 is automatically stirred and the cultured cells are automatically photographed at regular intervals by the microscope 300, the photograph becomes unclear due to wrinkles generated in the observation region on the observation hole 210, and the number of cells Count and cell state observation could not be performed properly.
The present invention solves such a problem by providing the cell culture container and / or the cell culture apparatus with means for preventing wrinkles.

[First embodiment]
Next, the cell culture container and cell culture method of the first embodiment of the present invention will be described with reference to FIGS. FIG. 4 is a diagram showing the configuration of the cell culture container of the present embodiment. FIG. 5 is a diagram showing the appearance and partial enlargement of the cell culture container of the present embodiment.
The cell culture container 10 of this embodiment is a container formed in a bag shape (back shape) using a soft wrapping material. By using a soft wrapping material as the material of the cell culture container 10, flexibility and flexibility can be imparted to the cell culture container 10. As the soft wrapping material, for example, those described in JP 2009-247225 A (culture container) and JP 2006-262676 A (culture bag, medium storage method and cell culture method) are used. Can do.
Moreover, the cell culture container 10 has gas permeability required for cell culture, and a part or all of the cell culture container 10 is transparent so that the contents can be confirmed. Examples of the culture vessel material that satisfies such conditions include polyolefins, ethylene-vinyl acetate copolymers, styrene elastomers, polyester thermoplastic elastomers, silicone thermoplastic elastomers, and silicone rubber.

  As described above, since the cell culture container 10 of the present embodiment is made of a soft packaging material, wrinkles are likely to occur. However, the cell culture container 10 is used as a fixing member 11 for preventing the generation of wrinkles. The frame 11a is provided. The frame 11a is adhered to the lower surface of the cell culture container 10, and as a result, a portion surrounded by the frame 11a on the surface of the cell culture container 10 is an area where generation of wrinkles is prevented.

  That is, the frame 11a is a rigid frame, and is adhered to the cell culture container 10 in a state where the surface of the cell culture container 10 is pulled to extend the ridge. In FIG. 4, a rectangular shape is illustrated, but the shape may be arbitrary. Further, the material of the frame 11a is not particularly limited. For example, polystyrene, polyethylene, polypropylene, PET, PMMA, cyclic polyolefin (COP), ABS, polyvinyl chloride, stainless steel, aluminum, glass, MC nylon, Engineering plastics such as polycarbonate can be used.

  The cell culture vessel 10 is filled with cells to be cultured and a culture solution. As the cultured cells, various cells in humans, other animals, plants, microorganisms and the like can be used. For example, cells in human leukemia T lymphoma, normal peripheral blood mononuclear cells, suspended human embryonic kidney cell lines, Hematopoietic stem cells and the like can be used. As the culture solution, those suitable for the growth of cultured cells can be used as appropriate, and are not particularly limited. For example, a liquid medium or a gel medium can be used.

  FIG. 5 shows a culture bag (photograph) manufactured as an example of such a cell culture container 10. A frame is adhered and fixed to the lower right portion of the culture bag. As shown in the partially enlarged view (photo), the surface of the culture bag surrounded by the frame is in a state where no wrinkles are generated due to tension. On the other hand, there are many wrinkles around the frame. It can be seen that fixing the frame to the observation region in the culture bag in this manner can prevent wrinkles from occurring in the observation region and improve the cell observation accuracy.

  The cell culture method of the present embodiment is a method in which cells and a culture solution are enclosed in such a cell culture vessel 10 and placed on a loading table to perform cell culture. At this time, at a predetermined timing, a stirring member such as a roller is moved in the horizontal direction while being pressed against the cell culture vessel 10, and the culture solution in the cell culture vessel 10 is stirred to make the culture solution uniform. it can. Thereby, it becomes possible to improve the proliferation efficiency of a cell.

Further, the cells in the cell culture container 10 are automatically photographed by the microscope 30 at regular intervals. In addition, although the imaging | photography of the cell in the cell culture container 10 by the microscope 30 is performed through the observation hole in the mounting base in which the cell culture container 10 is mounted, the mounting base is abbreviate | omitted in FIG.
Further, the microscope 30 is a microscope having a camera function. Although not shown, the captured image data is input to the information processing apparatus, and the number of cells in the image data is counted. As such a microscope 30, an automatic photographing method and a cell number counting method, those generally used at present can be used. The camera function may be provided separately from the microscope 30.

According to the cell culture method of this embodiment, since the cell culture container 10 is made of a soft wrapping material, wrinkles are likely to occur, but the observation area surrounded by the frame 11a is prevented from generating wrinkles. .
For this reason, according to the present embodiment, it is possible to clearly observe the number and state of cells by photographing the cells in the cell culture container 10 with the microscope 30 through this observation region. Yes.

[Second Embodiment]
Next, the cell culture container and cell culture method of the second embodiment of the present invention will be described with reference to FIG. This figure is a diagram showing the configuration of the cell culture container of the present embodiment.
The cell culture container 10 of the present embodiment includes a plate 11b as a fixing member 11 for preventing generation of wrinkles.
That is, the plate 11b is adhered to a part of the lower surface of the cell culture container 10 of the present embodiment, whereby the portion of the surface of the cell culture container 10 where the plate 11b is adhered This is an area where the problem is prevented.

The plate 11b is a flat plate that is transparent or rigid, all or part of which is capable of observing cells in the cell culture vessel 10 through the transparent portion. Examples of the material of the plate 11b include engineering plastics such as polystyrene, polyethylene, polypropylene, PET, PMMA, cyclic polyolefin (COP), ABS, polyvinyl chloride, silicone, and polycarbonate.
In addition, this plate 11b can also be set as the structure affixed on the whole lower surface of the cell culture container 10. FIG.

In addition, it is also preferable that the plate 11b is adhered to the upper surface above the observation region in the cell culture container 10 and provided. In this way, a region in which the generation of wrinkles is prevented can be formed on the upper surface of the cell culture container 10, so that the light emitted from the illumination device 40 is scattered or refracted irregularly, so that the amount of light is reduced. It is possible to prevent non-uniformity and improve the accuracy of a photograph taken by the microscope 30.
About the other point in the cell culture container 10 of this embodiment, it can be set as the thing similar to 1st embodiment. Further, the cell culture method of the present embodiment can be the same as that of the first embodiment except that the cell culture container 10 provided with such a plate 11b is used.

[Third embodiment]
Next, the cell culture container and cell culture method of the third embodiment of the present invention will be described with reference to FIG. This figure is a diagram showing the configuration of the cell culture container of the present embodiment.
The cell culture container 10 of the present embodiment includes a high-thickness portion 11c formed on a part of the surface so as to have a rigid thickness. The high-thickness portion 11c is formed by thickening a part of the soft wrapping material constituting the cell culture container 10 to form a rigid, transparent and flat observation region, and is a region where generation of wrinkles is prevented. is there.
In the present embodiment, the cells in the cell culture container 10 can be appropriately observed by taking an image with the microscope 30 through such an observation region.

In the present embodiment, it is also preferable to provide the high-thickness portion 11 c not only on the lower surface of the cell culture container 10 but also on the upper surface above the observation region in the cell culture container 10.
If it does in this way, it can prevent that the light irradiated from the illuminating device 40 scatters or refracts irregularly, and thereby it becomes possible to prevent the amount of light from becoming non-uniform, and to improve the accuracy of a photograph taken by the microscope 30. It becomes possible.

  About the other point in the cell culture container 10 of this embodiment, it can be set as the thing similar to 1st embodiment. The cell culture method of this embodiment can also be the same as that of the first embodiment except that the cell culture container 10 having such a high-thickness portion 11c is used.

[Fourth embodiment]
Next, a cell culture device and a cell culture method according to a fourth embodiment of the present invention will be described with reference to FIG. This figure is a diagram showing the configuration of the cell culture device of the present embodiment.
The cell culture device of this embodiment includes a loading table 20 on which a cell culture container is loaded.
The loading table 20 is a flat table on which the cell culture container is placed. The loading table 20 can be provided with various configurations for placing the cell culture container, such as a container fixture for fixing both ends of the cell culture container to the loading table 20. The loading table 20 includes an observation hole 21. Through this observation hole 21, the cells in the cell culture container loaded on the loading table 20 can be observed by the microscope 30.

The cell culture device of the present embodiment includes a fixing member 23 as a means for preventing wrinkles around the observation hole 21 of the loading table 20. As the fixing member 23, a rigid frame can be used. Further, as the fixing member 23, an adhesive member and / or a member formed of a material having a high friction coefficient, such as rubber, can be used.
As a cell culture container in this embodiment, a general thing can be used and the material can be the same as that of 1st embodiment. In the fifth to eighth embodiments relating to the cell culture apparatus provided with means for preventing the occurrence of wrinkles only on the loading platform 20, which will be described later, a common cell culture container should be used in the same manner. Can do.

  In the cell culture method of this embodiment, cells and a culture solution are sealed in a cell culture container and placed on the loading table 20 in this embodiment to perform cell culture. At this time, about the point that the culture solution in the cell culture vessel can be stirred and the culture solution can be homogenized, and that the cells in the cell culture vessel are automatically photographed by the microscope 30 at regular intervals, The same as in the first embodiment.

Here, in the present embodiment, the cell culture container is pressed against the loading table 20 provided with the fixing member 23, whereby tension is applied to a partial region on the lower surface of the cell culture container by the fixing member 23. And stretched to form a region where wrinkles are prevented.
Specifically, for example, a portion surrounded by the frame can be fixed without wrinkles by pressing the cell culture container against the frame-shaped fixing member 23 in a state where the cell culture container is stretched to the left and right. Further, the height of the fixing member 23 with respect to the loading table 20 is increased, for example, the thickness of the frame-shaped fixing member 23 is set to a size of 5 mm to 10 mm, and the cell culture container is pressed against the loading table 20. The part surrounded by the frame can be fixed without any wrinkles.
As described above, according to the present embodiment, the cells in the cell culture container are appropriately captured by photographing the cells in the cell culture container with the microscope 30 using the area in which the generation of wrinkles is prevented as the observation area. It can be observed.

[Fifth embodiment]
Next, the cell culture apparatus and cell culture method of 5th embodiment of this invention are demonstrated with reference to FIG. This figure is a diagram showing the configuration of the cell culture device of the present embodiment.
The cell culture device of this embodiment is different from that of the fourth embodiment in that the loading table 20 includes suction means 24 instead of the fixing member 23 as means for preventing wrinkles. About another point, it can be set as the thing similar to 4th embodiment.

The suction means 24 in the present embodiment includes a suction hole 24a on the side of the observation hole 21, and a negative pressure is applied to the surface of the cell culture container through the suction hole 24a. In the example of the figure, the suction holes 24a are provided at two places around the observation hole 21, but can be provided at three or four or more places.
When the cell culture container is placed on the loading table 20, the suction means 24 sucks the upper part around the observation hole 21 on the lower surface of the cell culture container. As a result, the observation region in the cell culture container can be stretched by applying tension, and wrinkles can be prevented from occurring in the region.
The suction means 24 is not particularly limited as long as it can suck the surface of the cell culture container and apply tension to the observation region to prevent generation of wrinkles.

In the cell culturing method of the present embodiment, a cell culturing apparatus provided with a suction means 24 on the loading table 20 is used. The cell culture container is placed on the loading table 20 to culture the cells, and the cells in the cell culture container are automatically photographed by the microscope 30 at regular intervals. In this embodiment, prior to photographing, the cells are cultured. The periphery of the observation area is sucked and stretched by the suction means 24.
Accordingly, even if wrinkles are generated in the observation region by, for example, stirring the cell culture container, it can be removed.

And after performing suction | inhalation in this way, imaging | photography of the cell in a cell culture container is performed with the microscope 30. FIG. It should be noted that the cells in the cell culture container can be photographed by the microscope 30 while performing suction by the suction means 24.
As described above, according to the cell culture device and the cell culture method of the present embodiment, since wrinkles are not reflected in the photograph taken by the microscope 30, it is possible to appropriately observe the cells.

[Sixth embodiment]
Next, the cell culture apparatus and cell culture method of 6th embodiment of this invention are demonstrated with reference to FIG. This figure is a diagram showing the configuration of the cell culture device of the present embodiment.
The cell culture device according to this embodiment is different from the fourth embodiment in that the loading table 20 includes not the fixing member 23 but a pressing means 25 as a means for preventing wrinkles. About another point, it can be set as the thing similar to 4th embodiment.
The pressing means 25 is a flat plate provided above the observation hole 21 in the loading table 20 so as to be movable up and down. Although not shown in the figure, the pressing means 25 is supported and moved up and down by providing a supporting member for the pressing means 25 and a driving means on the loading table 20.

  The flat plate constituting the pressing means 25 is preferably of a size larger than the area for observing cells in the cell culture container. If the pressing means 25 is configured in this way, the cell culture container is placed on the loading table 20, and the pressing means 25 is moved downward to sufficiently press the region for observing the cells in the cell culture container. Thus, it becomes possible to remove the soot.

In the cell culturing method of the present embodiment, a cell culturing device provided with a pressing means 25 on the loading table 20 is used.
Also in the cell culture method of the present embodiment, the cell culture container is placed on the loading table 20 and the cells are cultured, and the cells in the cell culture container are automatically photographed by the microscope 30 at regular intervals. In this embodiment, prior to imaging, first, the pressing unit 25 is moved downward, and the pressing unit 25 presses an area for observing cells in the cell culture container against the loading table 20.
As a result, the observation region in the cell culture container is stretched, and even if wrinkles are generated in the observation region by, for example, stirring the cell culture container, it can be removed.

Then, after pressing the cell culture container, the pressing means 25 is gently moved upward to return to the original position, and the cells in the cell culture container are photographed by the microscope 30.
As described above, according to the cell culture device and the cell culture method of the present embodiment, since wrinkles are not reflected in the photograph taken by the microscope 30, it is possible to appropriately observe the cells.

[Seventh embodiment]
Next, the cell culture apparatus and cell culture method of 7th embodiment of this invention are demonstrated with reference to FIG. This figure is a diagram showing the configuration of the cell culture device of the present embodiment.
The cell culture device of this embodiment is different from that of the fourth embodiment in that the loading table 20 includes not the fixing member 23 but a moving means 26 as a means for preventing wrinkles. About another point, it can be set as the thing similar to 4th embodiment.

The moving means 26 includes a fixing member 26a that contacts the surface of the cell culture container, and a driving means 26b for moving the fixing member 26a. As shown in FIG. Can be arranged. It is also possible to arrange them at three or more locations around the observation hole 21. The driving means 26b can be constituted by a motor and a ball screw, for example.
The fixing member 26a is composed of a member having adhesiveness and / or a member having a high coefficient of friction. When the cell culture container is placed on the loading table 20, the fixing member 26a contacts the surface of the cell culture container. When the driving unit 26b moves the moving unit 26 in the opposite direction with respect to the observation hole 21, the movement of the moving unit 26 is caused by the fixing member 26a being adhered to the surface of the cell culture container or by frictional force. At the same time, the observation region in the cell culture container is stretched.
In the example of FIG. 11, the cell culture container placed on the loading table 20 contacts the moving means 26 at two places on both sides of the observation area, and the observation means is stretched by the movement means 26 moving in opposite directions. It is supposed to be.

In the cell culturing method of the present embodiment, a cell culturing apparatus provided with a moving means 26 on the loading table 20 is used.
Also in the cell culture method of the present embodiment, the cell culture container is placed on the loading table 20 to culture the cells, and the cells in the cell culture container are automatically photographed by the microscope 30 at regular intervals. However, in this embodiment, the observation area is first stretched by the moving means 26 prior to photographing. Thereby, even if wrinkles are generated in the observation region by, for example, stirring the cell culture container, it is possible to remove the wrinkles.

Then, after the cells in the cell culture container are photographed by the microscope 30, the moving means 26 can be returned to the original position. At this time, after the moving means 26 after moving is moved downward and separated from the cell culture container, the moving means 26 is moved to the original position, so that the moving means 26 is moved to the original position without generating wrinkles in the observation region. It can be returned to the position. In addition, after returning the moving means 26 to the original position without generating wrinkles in the observation area in this way, it is also possible to photograph the cells in the cell culture container with the microscope 30.
As described above, even with the cell culture device and the cell culture method of the present embodiment, since wrinkles are not reflected in the photograph taken by the microscope 30, it is possible to appropriately observe the cells.

[Eighth embodiment]
Next, a cell culture device and a cell culture method according to an eighth embodiment of the present invention will be described with reference to FIG. This figure is a diagram showing the configuration of the cell culture device of the present embodiment.
The cell culture device of this embodiment is different from that of the fourth embodiment in that the loading table 20 includes not a fixing member 23 but a pulling means 27 as a means for preventing wrinkles. About another point, it can be set as the thing similar to 4th embodiment.

As the pulling means 27, for example, a clamp or the like can be used, and the observation region in the cell culture container can be extended by grasping both ends of the cell culture container and pulling in the opposite direction. Although not shown in the drawings, the cell culture apparatus of the present embodiment is provided with driving means for moving the pulling means 27 in the opposite direction. This driving means can be constituted by, for example, a ball screw or a cylinder.
In the present embodiment, the cell culture container can be stretched by fixing one of the pulling means 27 and moving only the other pulling means 27. In addition, the cell culture apparatus may be provided with three or four or more pulling means 27 to pull the cell culture container in more directions.

In the cell culturing method of the present embodiment, a cell culturing apparatus provided with a pulling means 27 on the loading table 20 is used.
Also in the cell culture method of the present embodiment, the cell culture container is placed on the loading table 20 to culture the cells, and the cells in the cell culture container are automatically photographed by the microscope 30 at regular intervals. However, in this embodiment, the observation area is first stretched by the pulling means 27 prior to photographing. Thereby, even if wrinkles are generated in the observation region by, for example, stirring the cell culture container, it is possible to remove the wrinkles.
As described above, even with the cell culture device and the cell culture method of the present embodiment, since wrinkles are not reflected in the photograph taken by the microscope 30, it is possible to appropriately observe the cells.

[Ninth embodiment]
Next, the cell culture system and cell culture method of 9th embodiment of this invention are demonstrated with reference to FIG. This figure is a diagram showing the configuration of the cell culture system of the present embodiment.
As shown in FIG. 13, the cell culture system of this embodiment includes a cell culture container 10 and a cell culture device including a loading table 20. In the cell culture system of the present embodiment, both the cell culture container 10 and the loading table 20 are provided with means for preventing the occurrence of wrinkles in the observation region in the cell culture container 10, and by combining them, It is possible to enlarge the observation area.

  That is, the cell culture container 10 in this embodiment is provided with the coupling members 11d having convex shapes at the four corners around the observation region on the lower surface. Further, at the four corners around the observation hole 21 of the loading table 20, there are provided coupling holes 28 that fit into the coupling member 11d. When the cell culture container 10 is placed on the loading table 20, the coupling members 11 d provided at the four corners around the observation region of the cell culture container 10 are connected to the coupling holes provided at the four corners around the observation hole 21 of the loading table 20. 28 respectively. As a result, it is possible to stretch the observation region by applying tension.

  In the example of FIG. 13, four connecting members 11d of the cell culture container 10 and four connecting holes 28 of the loading table 20 are provided, but three or five or more connecting holes may be provided. Moreover, in the example of the same figure, the coupling member 11d has a convex shape and is configured to fit into the coupling hole 28. However, the coupling member 11d of the cell culture vessel 10 is composed of a magnet, and is coupled to the loading table 20. You may make it provide the coupling member which consists of a metal couple | bonded with 11d or a magnet.

The cell culture system of the present embodiment includes means for preventing wrinkles in the observation region on both the cell culture container 10 and the loading table 20, and the observation region can be extended by combining them. Any other means may be used, and means other than those described above for preventing wrinkles may be used. For example, both the cell culture container 10 and the loading table 20 may be provided with adhesive means, and by combining them, the observation region can be extended. A tenth embodiment to be described later is also an application example of this embodiment.
Other points such as the material of the cell culture vessel 10 and the configuration of the microscope 30 can be the same as those in the first embodiment.

In the cell culture method of the present embodiment, when the cell culture container 10 enclosing the cells and the culture solution is placed on the loading table 20, the coupling members 11d provided at the four corners around the observation region of the cell culture container 10 are The observation area is extended by applying tension to the connection holes 28 provided at the four corners around the observation hole 21 of the loading table 20.
Therefore, for example, when a culture solution in the cell culture container 10 is stirred by pressing a roller or the like against the cell culture container 10 during cell culture, generation of wrinkles in the observation region can be prevented. For this reason, when the cells in the cell culture container 10 are automatically photographed periodically from the microscope 30 through the observation region, it is possible to appropriately observe the cells without reflection of wrinkles. .

[Tenth embodiment]
Next, a cell culture system and a cell culture method according to a tenth embodiment of the present invention will be described with reference to FIG. This figure is a diagram showing the configuration of the cell culture system of the present embodiment.
As shown in FIG. 14, the cell culture system of this embodiment includes a cell culture apparatus including a cell culture container 10 and a loading table 20.
The cell culture container 10 includes an ear member 11e for fixing the observation region to the loading table 20, and the loading table 20 includes a pulling means 29 for grasping and pulling the ear member 11e.

The ear member 11e is disposed on both sides of the observation region on the lower surface of the cell culture container 10. In the example of the figure, each is arranged in the short direction of the cell culture vessel 10, but can also be arranged in the long direction. Moreover, it is preferable that the length of the ear member 11e is equal to or longer than the length of the side of the observation region on each side.
The pulling means 29 are arranged on both sides of the observation hole 21 in the loading table 20, and when the cell culture container 10 is placed on the loading table 20, each of the pulling means 29 holds the ear member 11e and pulls it downward. . Specifically, the pulling means 29 may include an actuator 29a for grasping the ear member 11d and a cylinder 29b for moving the actuator 29a downward.

  Thus, according to the cell culture system of the present embodiment, the ear members 11e arranged on both sides of the observation region of the cell culture container 10 are pulled by the pulling means 29 arranged on both sides of the observation hole 21 of the loading table 20. Can be pulled down. For this reason, the observation region is stretched to prevent wrinkles. Other points such as the material of the cell culture vessel 10 and the configuration of the microscope 30 can be the same as those in the first embodiment.

In the cell culture method of this embodiment, when the cell culture container 10 enclosing the cells and the culture solution is placed on the loading table 20, the ear member 11 e of the cell culture container 10 is grasped by the pulling means 29 of the loading table 20. Can be made. Then, the cells are cultured, and the cells in the cell culture container are automatically photographed by the microscope 30 at regular time intervals. In this embodiment, the ear member 11e is moved downward by the pulling means 29 prior to photographing. The image can be taken after pulling and extending the observation area to remove wrinkles. In addition, cell culture can also be performed by fixing the ear member 11e while being pulled downward by the pulling means 29 in this manner.
As described above, even with the cell culture system and the cell culture method of the present embodiment, since wrinkles are not reflected in the photograph taken by the microscope 30, it is possible to appropriately observe the cells.

The present invention is not limited to the above-described embodiments and examples, and it goes without saying that various modifications can be made within the scope of the present invention.
For example, in the above-described embodiments and examples, the case of culturing cells has been described. However, for example, in the case where observation of other organisms such as microorganisms, tissues, organs, or inorganic substances is necessary, cell culture containers are used. It is possible to make appropriate changes such as preventing wrinkles from occurring.

  The present invention can be suitably used for culturing cells in large quantities using a cell culture vessel.

DESCRIPTION OF SYMBOLS 10 Cell culture container 11 Fixing member 11a Frame 11b Plate 11c High-thickness part 11d Bonding member 11e Ear member 12 １３ 13 Culture solution 14 Cell 20 Loading platform 21 Observation hole 22 Container fixture 23 Fixing member 24 Suction means 24a Suction hole 25 Press means 26 Movement Means 26a Fixing member 26b Driving means 27 Pulling means 28 Joint hole 29 Pulling means 29a Actuator 29b Cylinder 30 Microscope 40 Illumination device

Claims (15)

  A cell culture container made of a soft wrapping material for enclosing cells and culture medium and culturing cells, the cell culture container being equipped with means for preventing the generation of wrinkles and preventing the generation of wrinkles A cell culture container characterized in that a region in which wrinkles are prevented is formed in at least a part of the cell culture container.   The cell culture container according to claim 1, wherein the means for preventing wrinkles is a rigid frame or plate.   The cell culture container according to claim 1, wherein the means for preventing the generation of wrinkles is a portion of the cell culture container formed to have a rigid thickness. A cell culturing apparatus equipped with a loading table on which a cell culture vessel made of soft packaging material is placed for enclosing cells and culture medium and culturing cells,
Means for preventing generation of wrinkles in the cell culture container is provided on the loading table, and at least one of the cell culture containers is mounted by means for preventing the generation of wrinkles by placing the cell culture container on the loading table. A cell culture device characterized in that a region where wrinkles are prevented is formed in a part.   The region in which generation of wrinkles is prevented in the cell culture container by pressing the cell culture container against the means for preventing the generation of wrinkles, wherein the means for preventing the generation of wrinkles is a rigid frame or plate The cell culture device according to claim 4, wherein the cell culture device is formed.   The means for preventing the generation of wrinkles is formed of a material having a high adhesiveness and / or coefficient of friction, and the cell culture container is pressed against the means for preventing the generation of wrinkles, thereby generating wrinkles on the cell culture container. The cell culture device according to claim 4, wherein a region in which the formation is prevented is formed.   A suction means is provided on the loading platform, and the area for observing the cells is generated by sucking and fixing the periphery of the area for observing the cells in the cell culture container by the suction means. The cell culture device according to claim 4, wherein the cell culture device is formed in a region in which the formation is prevented.   The loading table is provided with pressing means, and the area for observing the cells in the cell culture container is pressed against the loading table by the pressing means to prevent the generation of wrinkles. The cell culture device according to claim 4, wherein the cell culture device is formed in the formed region.   The loading platform is provided with moving means, and the moving means extends the area for observing the cells in the cell culture container, thereby forming the area for observing the cells in the area where generation of wrinkles is prevented. The cell culture device according to claim 4, wherein   5. The pulling means is provided on the loading table, and the cell culture container is pulled in the horizontal direction by the pulling means to form a region in which the generation of wrinkles is prevented in the cell culture container. The cell culture device described. A cell culture system comprising a cell culture container made of a soft wrapping material for enclosing cells and a culture solution and culturing the cell, and a loading table on which the cell culture container is placed,
There are provided coupling means for coupling the cell culture container to the loading table by coupling them to the periphery of the region for observing the cells in the cell culture vessel and the periphery of the observation hole in the loading table, respectively. A cell culture system characterized in that the region for observing the cells is formed in a region where generation of wrinkles is prevented by bonding.   As the coupling means, an ear member is provided around a region for observing cells in the cell culture container, and a pulling means is provided around an observation hole in the loading table, and the ear member is attached by the tension means. The cell culture system according to claim 11, wherein the region for observing the cells is formed in a region where generation of wrinkles is prevented by pulling. A cell culture method for culturing cells by enclosing cells and a culture solution in a cell culture vessel made of soft packaging material,
The cell culture container is provided with means for preventing the generation of wrinkles, and an area in which the generation of wrinkles is prevented is formed in at least a part of the cell culture container by the means for preventing the generation of wrinkles. A cell culture method comprising observing cells in a cell culture vessel. A cell culture method for culturing cells by enclosing cells and a culture solution in a cell culture vessel made of soft packaging material,
The loading table includes means for preventing generation of wrinkles in the cell culture container, and the cell culture container is placed on the loading table, and at least part of the cell culture container is formed by the means for preventing generation of wrinkles. A cell culture method comprising: forming a region where wrinkles are prevented from being observed, and observing cells in the cell culture vessel through this region. A cell culture method for culturing cells by enclosing cells and a culture solution in a cell culture vessel made of soft packaging material,
A coupling means is provided for coupling the cell culture container to each other around the region for observing cells in the cell culture container and the observation hole in the loading table, and fixing the cell culture container to the loading table. Forming a region for observing the cells in a region where generation of wrinkles is prevented, and observing the cells in the cell culture container through the region. .