JPH1175821A - Biochemical reaction apparatus for culturing several series of cell and biochemical reaction - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a biochemical reaction apparatus and a biochemical reaction method capable of individually culturing and controlling each cell in a culture system, capable of subjecting several series of cells to simultaneous culture or cocultivation by commonly using a reaction solution and individually culturing and controlling each cell in such a culture system that a product of a kind of cell can become a substrate of other kind of cell or a physiologically active substance, especially in culture of animal cell or plant cell. SOLUTION: This biochemical reaction apparatus is equipped with reactors 1 and 2 for retaining different cells and a reaction solution preparing tank 3. The reactors 1 and 2 receives feed of a reaction solution 5 by a pump 4 from the reaction solution preparing tank 3 and the reaction solution discharged from each reactor is returned to the preparing tank 3 and discharged reaction solutions are mixed and the mixture is circulated between each reactor and the preparing tank 3. When it is desired to prevent the cell from moving from the reactor 1 to the preparing tank 3, a filter 8 is fitted in an inlet flow path 6 and/or outlet flow path 7. When it is desired to feed a specific gas and liquid to the reactor 2, a feed flow path 10 is formed in the inlet flow path 9.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biochemical reaction apparatus for culturing a plurality of types of cells by sharing a reaction solution and a biochemical reaction method using the same, and more particularly, to a reaction by one cell. The present invention relates to a biochemical reaction apparatus and a biochemical reaction method for causing a product to act on another cell to cause a reaction, and is particularly effective for substance conversion and substance production by a biochemical reaction.

[0002]

2. Description of the Related Art Conventionally, various reaction systems in which reactors containing a plurality of cells or physiologically active substances are connected in series have been used. In such a reaction system, for example, a reaction solution in which an anaerobic bacterium has acted is subsequently supplied to an aerobic reaction tank, or after reacting one kind of microorganism, the microorganism is filtered and used for the next biochemical reaction. Or that is being done.

However, in such a series reaction system, after performing one reaction, it is necessary to adjust the reaction solution so as to be suitable for the next biochemical reaction. For example, in the reaction solution after the aerobic reaction,
Since dissolved oxygen and the substrate are consumed, there is a problem that it is necessary to replenish the oxygen and the substrate before the next aerobic reaction.

On the other hand, in the case of culturing animal cells, there has been reported an example in which a plurality of types of cells are mixed and cultured to produce a useful substance. For example, it has been reported that in a pure culture of liver parenchymal cells, the ability to produce albumin decreases in a few days, but when co-cultured with stellate cells, the ability to produce albumin could be maintained for one month or more.

[0005] However, when such co-culture is performed, there is a problem that rapidly growing cells predominantly proliferate, and it is difficult to control the distribution of cells as desired. This is remarkable in a co-culture system of different microorganisms.

The present invention has been made in view of the above-mentioned problems of the prior art, and an object of the present invention is to allow a plurality of types of cells to be co-cultured or co-cultured by sharing a reaction solution. In particular, in the cultivation of animal cells and plant cells, a biochemical reaction device capable of individually culturing and managing each cell in a culture system in which a product of one cell can be a substrate or a physiologically active factor of another cell, and It is to provide a biochemical reaction method.

[0007]

Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventors have connected a reaction solution adjusting tank and each reactor containing separate cells or cell groups in parallel, The present inventors have found that the above object can be achieved by adopting a structure in which the reaction solution discharged from each reactor returns to the reaction solution adjustment tank, and have completed the present invention.

That is, the biochemical reaction apparatus of the present invention comprises a plurality of reactors for individually accommodating a plurality of types of cells in one kind or a combination of two or more types, and a reaction solution containing a reaction solution to be supplied to the cells. An adjustment tank, and the reaction solution adjustment tank sends the reaction solution to the plurality of reactors in parallel,
When the reaction liquid discharged from these reactors returns to the reaction liquid adjustment tank, the reaction liquid circulates between the reaction liquid adjustment tank and each reactor, and during this circulation, the reaction liquids discharged from the reactor are separated from each other. The cells are merged, and the plurality of types of cells share a reaction solution.

[0009] In a preferred embodiment of the biochemical reaction device of the present invention, the cells in at least one of the above reactors are:
It is characterized by comprising a movement preventing means for preventing movement to another reactor.

Further, another preferred embodiment of the biochemical reaction apparatus of the present invention is characterized in that the cells are animal and / or plant cells, and in particular, the animal cells are two or more kinds of cells constituting a liver. In the case of, a remarkable effect of simultaneous culture or co-culture is obtained.

Further, a biochemical reaction method of the present invention is characterized by using the above-described biochemical reaction device.

[0012]

According to the biochemical reaction apparatus and method of the present invention, cells can perform independent biochemical reactions in a plurality of reactors using a common reaction solution or culture solution. In addition, a product of a biochemical reaction in one reactor can be easily supplied to another biochemical reaction.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. As described above, the biochemical reaction device of the present invention includes a plurality of reactors that individually store a plurality of types of cells, and a reaction solution adjustment tank that stores a reaction solution to be supplied to these cells.

Here, it is sufficient that the reactor can accommodate the cells to be cultured. One reactor may accommodate one type of cells, but a plurality of types of cells may be accommodated.
It is also possible to accommodate in one reactor. Further, the cells to be cultured by the present invention are microorganisms, plant and animal cells, and as a reactor for accommodating them, a culture device containing cells, a container containing a carrier on which cells are fixed, a cell, A so-called hollow fiber type bioreactor that supplies gas and a substrate through a hollow fiber arranged in this space and a combination thereof can be used.

It is sufficient that the reaction solution adjusting tank can contain a reaction solution involved in causing various kinds of biochemical reactions of target cells. The reaction solution includes a substrate, a reaction inhibitor and a physiologically active substance. And the like. When the reaction solution is consumed in culturing or the like, or the components change, the reaction solution may be supplied to the reaction solution adjusting tank, or a part of the reaction solution may be withdrawn and supplied.

In the present invention, the reaction solution in the reaction solution adjusting tank is sent to a plurality of reactors in parallel, and the reaction solution discharged from these reactors returns to the adjusting tank. That is, the reaction solution circulates between the reaction solution adjustment tank and each reactor, and the reaction solution discharged from each reactor after the reaction (in some cases, the reaction solution containing the reaction product) is discharged from the reaction solution. Until the adjustment tank, the reaction liquids typically join in the reaction liquid adjustment tank, and the discharged reaction liquids are mixed.

After the reaction liquids are combined as described above, the reaction liquid is sent to each reactor again, so that the cells in each reactor share the reaction liquid, and the cells share the reaction liquid. When co-cultured or co-cultured, if a certain reactor cell produces a reaction product, this reaction product is mixed into the reaction solution and subjected to a biochemical reaction by another reactor cell. Will be.

The method of sending the reaction solution to each reactor includes a method of discharging the reaction solution by a pump and then branching and sending the reaction solution to the reactor, a method of providing a dedicated pump in each reactor to send the reaction solution to the reactor, and a method using a pump instead of the pump. Various methods can be selected, such as a method of feeding under pressure such as hydrostatic pressure or a combination thereof. When it is desired to accurately control the supply speed of the reaction solution to each reactor, a dedicated flow rate adjusting mechanism may be provided for each reactor.

Further, in the biochemical reaction device of the present invention,
If it is necessary to prevent cells of a certain reactor from migrating to another reactor, a means for preventing migration may be provided. This movement preventing means may be provided upstream of the reactor to prevent inflow or downstream of the reactor to prevent outflow. Specifically, a filter for capturing cells, a heat sterilizer, ultraviolet light, etc. Optical sterilizer and radiation sterilizer.

As described above, in the present invention, cells such as microorganisms, plants, and animals can be used as cells to be housed and held in the reactor and cultured. In particular, many plant and animal cells have a slower growth rate than microorganisms. Therefore, a reactor containing the same is easy to control, and is effective for performing a biochemical reaction while controlling each reactor.

Among animal cells, the cells constituting the liver perform biochemical reactions while a plurality of types of differentiated cells interact with each other, and the present invention makes it easy to develop an in vitro model of such a system. It will be realized. That is, in a living body, these cells are in almost the same environment while being in contact with each other. However, according to the biochemical reaction device of the present invention, the supply of a common reaction solution or culture solution from the reaction solution adjusting tank causes It is possible to carry out almost the same biochemical reaction and cultivation. Specifically, the parenchymal cells, stellate cells, fibroblasts, etc. constituting the liver are cultured in respective reactors, and a common reaction solution or culture solution may be sent to each reactor from the above-described reaction solution adjustment tank. become.

The biochemical reaction method of the present invention uses the above-described biochemical reaction apparatus of the present invention, and includes a decomposition reaction, a synthesis reaction of proteins and nucleic acids, a substance conversion reaction, and a combination thereof. Applicable to various biochemical reactions. For example, albumin can be produced continuously by culturing liver real cells and stellate cells in separate reactors, and culturing them while circulating a common culture solution through a reaction solution adjusting tank. The present invention is particularly effective for such biochemical reactions.

[0023]

Hereinafter, the present invention will be described in more detail with reference to the drawings, but the present invention is not limited to these embodiments.

(Embodiment 1) FIG. 1 is a block diagram showing one embodiment of the biochemical reaction apparatus of the present invention. In this figure, this biochemical reaction apparatus includes reactors 1 and 2 that house and hold different cells, and a reaction solution adjusting tank 3. In the apparatus of the present embodiment, the reactors 1 and 2 receive the supply of the reaction liquid 5 from the reaction liquid adjustment tank 3 by the pump 4, and the reaction liquid discharged from each reactor returns to the adjustment tank 3, and the discharged reaction liquid is returned to the adjustment tank 3. The liquids are mixed and circulated again between each reactor and the adjustment tank 3.

When it is desired to prevent cells from moving from the reactor 1 to the regulating tank 3 (ie, to prevent the cells from flowing into the reactor 2), the inlet channel 6 and / or Alternatively, a filter 8 for preventing passage of cells may be provided in the outlet channel 7. When it is desired to supply a specific gas or liquid to the reactor 2, a supply channel 10 can be provided in the inlet channel 9 of the reactor 2.

Next, an example of a biochemical reaction method using the apparatus of this embodiment will be described. First, reactors 1 and 2
Is filled with a porous carrier serving as a scaffold for cell adhesion, for example, porous glass, cellulose or porous gelatin beads, and the like. Glue and hold. Then, a culture solution 5, which is an example of a reaction solution filled in the adjustment tank 3, is sent out to each actor by the pump 4, and the culture solution 5 may be used to culture the parenchymal cells and the stellate cells. In this case, in the adjusting tank 3, it is preferable to appropriately adjust dissolved oxygen, pH and various substrate concentrations of the culture solution, and to stir the mixture so as to be uniform.

In such a reaction system, each cell proliferates without being inhibited by other cells, so that a desired number of cells can be secured. Further, according to this reaction system, it is possible to realize a reaction system in which the product of one cell continuously affects the production of protein of another cell, as in a living body. As the cells, not only animal cells such as liver cells, but also plant cells, microorganisms, and combinations thereof can be used.

(Embodiment 2) FIG. 2 is a block diagram showing another embodiment of the biochemical reaction apparatus of the present invention. Two types of microorganisms are cultured in separate culture tanks 21 and 22, respectively, and a substrate tank is provided. 23 shows a reaction system sharing 23. As described above, different microorganisms are cultured in the culture tanks 21 and 22,
The substrate tank 23 and the culture tanks 21 and 22 are connected by flow paths 24, 25, 26, and 27 provided with the pump 4, and the respective culture tanks 21 and 22 and the substrate tank form a circulation path. I have.

A heat exchange pipe 28 is provided in the heat exchange chamber 29 in the middle of the flow path 26 from the culture tank 22 to the substrate tank 23, and steam enters and exits the heat exchange chamber 29. The culture solution in the exchange pipe 28 can be heated and sterilized.

Next, an example of a biochemical reaction method using the apparatus of this embodiment will be described. While lactic acid bacteria are cultured in the culture tank 21 and lactic acid fermentation is performed by the lactic acid bacteria, methane bacteria are cultured in the culture tank 22 and methane fermentation is performed. Here, Japanese Patent No. 1902791 describes that when a liquid obtained by heat-treating a culture solution containing methane bacteria is added to a methane fermentation solution, methane fermentation is promoted. It has been found that a liquid obtained by heat-treating a liquid containing bacteria promotes lactic acid production by lactic acid bacteria and mold. This phenomenon suggests that the solution obtained by heat treatment of methane bacteria also promotes the physiological activity of other bacteria other than methane bacteria. In the reaction system of the present example, a liquid obtained by heat-treating a liquid containing methane bacteria promotes lactic acid fermentation and methane fermentation. In addition, there is an advantage that the microorganisms in the culture solution 22 do not move to the culture tank 21 by the heat treatment in the heat exchange chamber 29.

[0031]

As described above, according to the present invention, the reaction solution adjusting tank is connected in parallel with each reactor containing a separate cell or group of cells, and the reaction solution discharged from each reactor is Since it is configured to return to the reaction solution adjusting tank, a plurality of types of cells can be co-cultured or co-cultured by sharing the reaction solution. A biochemical reaction device and a biochemical reaction method capable of individually culturing and managing each cell in a culture system that can serve as a substrate or a bioactive factor for other types of cells can be provided.

In the present invention, since the reaction solution is sent from a common reaction solution adjusting tank to a plurality of types of cells, not only the number of adjusting tanks is reduced but also the complexity of adjusting the culture solution is reduced. It is reduced. Further, the products of one reaction can be subjected to the reaction of another cell. This product may be a substrate for other cells or a physiologically active substance.

[Brief description of the drawings]

FIG. 1 is a configuration diagram showing one embodiment of a biochemical reaction device of the present invention.

FIG. 2 is a configuration diagram showing another embodiment of the biochemical reaction device of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Reactor 2 Reactor 3 Adjustment tank 4 Pump 5 Reaction liquid 6 Inlet flow path 7 Outlet flow path 8 Filter 9 Inlet flow path 10 Supply flow path 21 Reactor 22 Reactor 23 Substrate tank 24, 25, 26, 27 Flow path 28 Heat exchange pipe 29 heat exchange room

Claims (5)

[Claims] 1. A reactor comprising: a plurality of reactors for individually accommodating a plurality of kinds of cells in one kind or a combination of two or more kinds; and a reaction solution adjusting tank containing a reaction solution to be supplied to the cells. The adjustment tank sends the reaction solution to the plurality of reactors in parallel, and the reaction solution discharged from these reactors returns to the reaction solution adjustment tank, so that the reaction solution flows between the reaction solution adjustment tank and each reactor. A biochemical reaction device, wherein the reaction solution discharged from the reactor merges during the circulation, and the plurality of types of cells share the reaction solution. 2. The biochemical reaction device according to claim 1, further comprising a movement preventing means for preventing cells in at least one of the reactors from moving to another reactor. 3. The biochemical reaction device according to claim 1, wherein the cells are animal and / or plant cells. 4. The biochemical reaction device according to claim 3, wherein the animal cells are two or more types of cells constituting a liver. 5. A biochemical reaction method using the biochemical reaction device according to any one of claims 1 to 5.