JP4534105B2 - Composite cell culture apparatus and culture method - Google Patents

Description

[0001]
[Industrial application fields]
The present invention relates to a composite cell culture apparatus and a culture method for culturing different culture systems while supplying cells to each system while holding the cells in each system or in effect, to microorganisms, animal cells, It is used for searching plant cells, searching for biochemical substances, searching for culture products, and the like.
[0002]
[Background Art and Problems to be Solved by the Invention]
It is said that 10% or less of all microorganisms can be cultured by conventional techniques. The microorganisms that cannot be cultured include symbiotic microorganisms that cannot grow unless they coexist with other microorganisms. When microorganisms are symbiotic in a nutrient-rich culture solution, only the dominant microorganisms will grow, so the microorganisms will not pass for the cultivation of the symbiotic microorganisms, but the culture solution components will pass through the diaphragm, and the culture medium will be sandwiched between them. A co-cultivation apparatus for culturing different microorganisms in each of the opposing culture systems is used.
[0003]
However, in the above symbiotic culture device, parts and a special culture device are required to form a sealed structure so that microorganisms do not pass by partitioning the culture system with a diaphragm, so the culture device is also expensive and must be large, Small and simple culture vessels such as conventionally used test tubes and Erlenmeyer flasks cannot be used.
[0004]
It is known that heterogeneous cells coexist in vivo in animals, but it is difficult to coexist with cells in a balanced manner outside the body, and its general method has not been established. .
[0005]
Although the inhibition circle method is used as a method for evaluating the titer of antibiotics, etc., it was necessary to use the inhibition circle method after culturing and separating and purifying antibiotics.
[0006]
A composite culture system in which a porous carrier such as urethane or cellulose sponge is introduced into a culture solution and anaerobic culture is carried out inside the carrier and aerobic culture outside the carrier and on the surface of the carrier is used for activated sludge treatment, etc. Since cells can move between the inside and outside of the carrier, it is impossible to construct an aerobic or both anaerobic culture system while holding different types of cells. Furthermore, it was difficult to take out the cells in the porous carrier, and there was inconvenience when collecting the cells.
[0007]
The problem to be solved by the present invention is to provide a complex cell culture apparatus and a culture method that can be realized even with simple culture instruments such as test tubes and flasks in view of the above-mentioned conventional drawbacks.
[0008]
[Means for Solving the Problems]
While examining the above problems, the present inventors have found that a complex cell culture system can be easily realized by arranging a liquid-permeable tube containing cells and culture medium in another culture system, and completed the present invention. .
[0009]
That is, the present invention is a composite cell culture apparatus, in which a tube containing a cell and a culture solution and having openings at both ends hermetically sealed and capable of passing the culture solution is disposed in a culture solution containing cells in a culture vessel. It is characterized by. The preferable form is characterized in that at least one cell inside and outside the tube does not pass through the tube. Furthermore, one of the preferable forms is characterized in that the inner diameter of the liquid-permeable tube does not exceed 2 mm.
[0010]
The present invention also relates to composite cell culture method, characterized that you culturing cells using the composite cell culture device.
[0011]
[Action]
By arranging a tube that can pass through the culture solution containing the cells and the culture solution in the culture solution containing the cells, the culture solution components can be moved in and out of the tube, and a complex culture system can be formed. . A complex culture system is a culture system in which two or more different culture systems have other positive or negative influences. Even if the metabolism is different, the same cells can be said to be different culture systems. This is because even in the same cell, different culture systems with different metabolism may be formed due to different culture solution compositions and dissolved gas concentrations. Therefore, the cells inside and outside the tube can be the same or different. In the present invention, it is necessary to retain the cells and the culture solution to such an extent that the cells maintain different culture systems inside and outside the tube even if they move between the culture systems.
[0012]
By not allowing at least one of the cells to pass through the tube, the tube can maintain a symbiotic balance without affecting the other cells. For example, a complex culture system can be maintained if cells in one culture system cannot be proliferated even if they move to the other culture system. It is desirable to select the permeability of the tube according to the size and molecular weight of the object that the cell, virus, protein, etc. wants to pass through or is prevented from passing through. If the inner diameter of the tube exceeds 2mm, aerobic cells grow in the tube. Even if dissolved oxygen is present outside the tube, the dissolved oxygen does not diffuse to the center of the tube and the inside becomes anaerobic. There are cases where it is not preferable when trying to keep in mind.
[0013]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described. As the first culture system, commonly used culture equipment such as test tubes, various flasks, and culture tanks can be used. A liquid-permeable tube containing a cell and a second culture system containing the culture solution is placed in the culture solution. As the tube, a filtration membrane, a dialysis membrane, an ion exchange membrane, a reverse osmosis membrane or the like can be used according to the purpose. The tube needs to store and hold the cells and the culture solution as long as the cells do not affect the culture even if the cells move through the culture system.
[0014]
When the first culture system and the second culture system have different cell types, it is desirable that at least one of the cells does not pass through the tube. If cells move in and out of the tube, only the dominant cells may grow. As long as it is possible to distinguish between both anaerobic bacteria in the tube and aerobic bacteria outside the tube, the cells may pass through the tube in both directions.
[0015]
【Example】
The present invention will be described with reference to examples. FIG. 1 shows a cross-sectional view of an embodiment of the present invention. A first culture system containing a culture solution 2 and microorganisms is formed in a test tube 1 arranged obliquely. The opening of the test tube 1 is plugged with a breathable cotton plug 3, which is shaken with a shaking device (not shown) and cultured aerobically. Inside the test tube, the microorganism and the culture solution forming the second culture system are enclosed in a hydrophilic tube 4 having an inner diameter of 1 mm formed of a porous cellulose membrane having a pore diameter of 0.2 μm or less, and the openings at both ends are plugged. 5 is sealed.
[0016]
Embodiments of the Examples
In the culture system of this example, the first and second culture systems can be aerobically cultured. A culture system can also be formed by a combination of microorganisms, plants and animal cells. It can be used both in the case where each culture system or one culture system exerts a positive effect on growth and substance production and on the other when it exerts a negative effect.
[0017]
Although the present invention has been described with reference to examples, the present invention is not limited to the examples.
For example, in the embodiment, a stopper is inserted into the opening of the tube to seal the tube, but the tube may be crushed or may be sealed by heat sealing according to the material of the tube. Moreover, it is desirable to sterilize the cells adhering to the outer wall or terminal of the tube with a flame or a chemical solution.
[0018]
【The invention's effect】
According to the present invention, cells can be complex-cultured using conventional culture equipment.
[Brief description of the drawings]
FIG. 1 shows a cross-sectional view of an embodiment of the present invention.
[Explanation of symbols]
1 Test tube 2 Culture solution 3 Cotton plug 4 Tube 5 plug

Claims (4)

An apparatus for cultivating a composite cell, comprising: a tube containing a cell and a culture solution and having a closed opening at both ends thereof and capable of passing the culture solution in a culture solution containing cells in a culture vessel . 2. The complex cell culture apparatus according to claim 1, wherein the tube does not allow passage of at least one cell inside and outside the tube . The composite cell culture apparatus according to claim 1 or 2, wherein an inner diameter of the tube does not exceed 2 mm . How complex cell culture using the composite cell culture apparatus characterized that you culturing cells according to claim 1.

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