JPH0779770A - Cell culture of organisms and culture tank therefor - Google Patents

Abstract

PURPOSE:To accelerate the proliferation of organic cells and enable a large volume of cell culture by feeding the culture medium to the cell inoculated on the tray as the medium is foamed and raised up to the cell from below the tray in the culture tank. CONSTITUTION:In the culture tank 1, the organic cells are placed on the tray 11 and the culture medium is stored in the bottom lower than the tray 11. The culture medium is foamed by means of the bubbler 15 at the bottom and the foam is grown and raised so that foamed medium is fed to the cells.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for culturing biological cells and a culturing apparatus, and more particularly to a method for promoting the culturing of biological cells by bringing them into contact with a foam culture medium and a large-scale culturing. The present invention relates to a culture device suitable for.

[0002]

2. Description of the Related Art In the following description, a plant tissue will be described as a representative example, but this does not exclude the case of targeting biological cells in general. When culturing plant tissue, the medium is supplied by the following method. (1) A method of solidifying a culture medium with agar or the like to prepare a solid medium. (2) A method of supplying a culture solution through a nozzle provided on the upper part of the culture tank.

However, all of these methods are unsuitable for the case where cells proliferate into a large cell mass or large-scale culture of plant cells using a large apparatus. For example, in the method using the solid medium, it is practically difficult to increase the size of the culture device, and in the method using the nozzle,
Since the amount and range (spreading) of the culture solution supplied from each nozzle is limited, when the cells grow into a large cell mass, the culture solution should be uniformly supplied to the entire cell mass. Will be difficult. Therefore, it is necessary to use a large-scale culture device equipped with a large number of nozzles, and there is a problem that the structure is complicated and the operation becomes complicated.

[0004]

The present invention has been made to solve the above problems, and an object thereof is to provide a culturing method capable of easily culturing a large amount of biological cells, and a simple structure. An object of the present invention is to provide a culture device that has and is suitable for mass culture.

[0005]

Means for Solving the Problems The culturing method of the present invention is carried out by contacting biological cells inoculated on a plate in a culture tank with a culture solution below the plate being raised in a foam shape and brought into contact therewith. The gist is to achieve the growth of the biological cells. In a preferred embodiment, in the above method, the foam culture medium contacting with the biological cells is once dipped, and then the cycle of contacting the foam culture medium with the biological cells again is repeated to grow the biological cells. The apparatus for carrying out the above method is not particularly limited, but in the present invention, the following 2
Present one device.

[0006] First, the first culture device has, in a culture tank, one to a plurality of trays on which biological cells are placed, and a foamer for foaming a culture solution contained below the trays. It has the gist.

The second culturing apparatus is provided with a culturing tank and a bubble forming tank, and the former culturing tank mounts biological cells.
~ Having a plurality of trays, the latter foam forming tank has a whisk for foaming the culture solution, between these culture tank and foam forming tank,
The gist of the present invention is to have a foam feeding pipe for feeding the foamy culture liquid to the culture tank and a liquid feeding pipe for feeding the culture liquid accumulated in the lower portion of the culture tank to the foam forming tank.

[0008]

Actions and Examples The culturing method of the present invention is characterized in that a foamy culture solution grown and raised from under the tray is brought into contact with the biological cells on the tray. And in a particularly preferred embodiment, the cycle in which the foam culture medium contacting the biological cells is once stopped and then the foam culture medium is again contacted with the biological cells is repeated, whereby the biological cells are always kept fresh. Since the culture medium can be supplied, the growth of biological cells can be further promoted.

Hereinafter, the culturing method of the present invention will be described in detail by taking the case of using the above-mentioned first or second culturing apparatus as an example, but these apparatuses and methods are for showing representative examples. There is no intention to limit the present invention. First,
The culture device of the present invention will be described in detail.

FIG. 1 shows the first culture device of the present invention.
It represents a culture device having one tray. The culture tank 1 includes a sterilizer and a temperature controller 10. In the upper part of the culture tank 1, an implantation recovery port 12 for inoculating biological cells and recovering after culturing, and a ventilation port 13 are provided. Inside the culture tank 1, a shelf 11 for placing biological cells is installed. Such a shelf is
It is not particularly limited as long as it is one that is usually used for culturing biological cells and can mount biological cells,
As such an example, for example, a perforated plate or a grooved plate can be cited. Below the culture tank 1, the culture solution is stored to a position lower than the height of the tray 11. A foamer 15 is installed at the bottom of the culture tank, and the culture solution is formed into foam by ventilating from the foamer vent 14 and further grown and raised one after another. The foaming device used in the present invention is not particularly limited, and any device can be used as long as it can grow and raise the culture solution in a foam shape, and examples thereof include a sparger and the like.

FIG. 2 shows the first culture device of the present invention.
1 illustrates a culture device having a plurality of shelves. When culturing a large amount of biological cells, an apparatus with a plurality of shelves provided in this manner is suitable for use, and a foaming device is used to facilitate the formation of a foam culture solution in the entire culture tank. Can be supplied to. The culture tank 1 is equipped with a sterilizer and a temperature controller (not shown). Inside the culture tank 1, a plurality of trays 11 for arranging biological cells are arranged, and on the side surface of the culture tank 1, biological cells are inoculated in each tray, and collected after culturing. An implantation recovery port 12 is provided for this purpose. Further, a ventilation port 13 is provided on the upper and side surfaces of the culture tank 1. At the bottom of the culture tank 1, a liquid reservoir 16 for storing the culture liquid is provided. A foamer 15 is installed at the bottom of the liquid reservoir, and the culture solution is foamed by ventilating from the foamer vent 14.

Next, FIG. 3 shows a culturing apparatus having a plurality of shelves in the second culturing apparatus of the present invention. The culture device comprises a culture tank 1 and a foam forming tank 2, and each of these tanks is equipped with a sterilizer and a temperature controller (not shown). This culture device is provided with a plurality of trays like the first culture device of the present invention shown in FIG. 2, but by providing the culture tank 1 and the foam forming tank 2 separately, The formation efficiency is enhanced, and it is also effective to install a stirring blade in the foam formation layer 2. Inside the culture tank 1, a plurality of trays 11 for placing biological cells are arranged, and on the side surface of the culture tank 1, biological cells are inoculated for each tray, and after culturing. Implantation recovery port 1 for recovery
Two are provided. A ventilation port 13 is provided on the upper and side surfaces of the culture tank 1. The culture medium is stored in the foam forming tank 2, and a foaming device 22 is installed at the bottom of the foam forming tank 2. The culture liquid grows and rises in a foam shape by ventilating from the foaming device ventilation port 21. Let Culture tank 1 and bubble formation tank 2
Are connected by a bubble feeding pipe 3 and a liquid feeding pipe 5, and in detail, the liquid feeding pipe 5 is connected via a liquid feeding pump 4. Further, the foam tube 3 can be connected to each shelf,
As a result, the culture solution can be evenly distributed over the entire biological cells. Next, a method for culturing plant tissue using each of the above-mentioned culture devices will be described in detail.

FIG. 4 illustrates a process of culturing plant tissue using the culturing apparatus of FIG. (A) After sterilizing the culture tank 1 in advance, the tray 11 is inoculated with the plant tissue as a seed (1). (B) Aeration is performed through the foamer vent 14 so that the culture solution that has been sterilized in advance in the culture tank 1 is foamed by the foamer 15 (2). (C) Aeration is performed until the foam culture solution covers the entire plant tissue (3). (D) Stop ventilation (4). (E) The bubbles burst and the volume of the foamy culture solution decreases to a position lower than the tray 11 (5). (F) Steps (b) to (e) are repeated. During this time, ventilation is performed from the ventilation outlet 13. By repeating this step, it is possible to constantly bring the fresh culture solution into contact with the plant tissue. (G) Plant tissue grows (6).

In this culturing step, from the start of aeration until the foamy culture solution covers the entire plant tissue [(b)-
The time required for (c)] is about 1 minute, and until the volume of the foamy culture medium decreases after the ventilation is stopped [(d)-
The time required for (e)] is about 3 minutes.

FIG. 5 illustrates a process of culturing plant tissue using the culturing apparatus of FIG. (A) The culture tank 1 is sterilized in advance, and a plurality of trays 11 are inoculated with plant tissue as a seed (1). (B) Aeration is performed through the foamer vent 14 so that the culture solution that has been sterilized in advance in the culture tank 1 is foamed by the foamer 15 (2). (C) Aeration is performed until the foam culture solution covers the entire plant tissue (3). (D) Stop ventilation (4). (E) The bubbles burst to reduce the volume of the foam culture medium.
The burst culture liquid returns to the liquid reservoir 16 (5). (F) The volume of the above-mentioned foamy culture solution is the ventilation port 1 on the side of the culture tank.
At the time when the gas has decreased from 3 to the extent that it does not blow out, ventilation is performed from the ventilation outlet 13 [(6) and (7)]. This is for defoaming the foamy culture solution attached to the plant tissue by performing aeration. This aeration is carried out until the foam culture medium is completely defoamed and the moistness in the plant tissue disappears. (G) Repeat steps (b) to (f). By repeating this process, it is possible to constantly bring fresh culture medium and air into contact with the plant tissue. (H) Plant tissue grows (8).

In this culturing step, from the start of aeration until the foamy culture solution covers the entire plant tissue [(b)-
The time required for (c)] is about 3 minutes, and until the volume of the foamy culture medium decreases after the ventilation is stopped [(d)-
The time required for (f)] is about 5 minutes.

FIG. 6 illustrates a plant tissue culturing process using the culturing apparatus of FIG. . (A) The culture tank 1 is sterilized in advance, and a plurality of trays 11 are inoculated with plant tissue as a seed (1). (B) Aeration is performed from the bubbler vent 21 so that the previously sterilized culture solution in the culture tank 1 is foamed by the bubbler 22. The foamy culture solution is sent to the culture tank 1 through the foaming tube 3 (2). (C) Aeration is performed until the foam culture solution covers the entire plant tissue (3). (D) Stop ventilation (4). (E) The bubbles burst to reduce the volume of the foam culture medium.
The burst culture solution is stored in the lower part of the culture tank 1, and the collected culture solution is sent to the bubble forming tank 2 through the solution sending pipe 5 and the solution sending pump (5). (F) When the volume of the foamy culture solution has decreased to the extent that it does not blow out from the ventilation outlet 13, ventilation is performed from the ventilation outlet 13 on the side of the culture tank [(6) and (7)]. This is for defoaming the foamy culture solution attached to the plant tissue by performing aeration. This aeration is carried out until the foam culture medium is completely defoamed and the moistness in the plant tissue disappears. (G) Repeat steps (b) to (f). By repeating this process, it is possible to constantly bring fresh culture medium and air into contact with the plant tissue. (H) Plant tissue grows (8).

In this culture step, aeration is started until the foam culture medium covers the entire plant tissue [(b) to (c)].
It takes about 3 minutes, and the time required from [(d) to (f)] after stopping aeration until the volume of the foamy culture solution decreases is about 5 minutes.

[0019]

EFFECTS OF THE INVENTION In the culture method of the present invention, since the foamy culture solution grown and raised from the bottom of the tray is brought into contact with the biological cells on the tray, the cells proliferate into a large cell mass. Or
Even when a large amount of cells are cultured using a large apparatus, the culture solution can be uniformly and easily supplied to the entire cells. The culture device of the present invention has a simple structure including a foamer and one to a plurality of shelves, and is suitable for large-scale culture of biological cells.

[Brief description of drawings]

FIG. 1 is a schematic view showing an example of a first culture device of the present invention.

FIG. 2 is a schematic diagram showing another example of the first culture device of the present invention.

FIG. 3 is a schematic view showing an example of a second culture device of the present invention.

FIG. 4 is a schematic diagram of a plant cell culturing process using the first culturing apparatus of the present invention.

FIG. 5 is a diagram schematically illustrating a plant cell culturing process using another first culturing apparatus of the present invention.

FIG. 6 is a schematic diagram of a plant cell culturing process using the second culturing apparatus of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Culture tank 2 Foam formation tank 3 Foaming pipe 4 Foaming pump 5 Liquid feeding pipe 10 Sterilizer and temperature control device 11 Perforated plate or groove plate 12 Implant recovery port 13 Passage exhaust port 14, 21 Foamer vent port 15, 22 Foamer 16 Liquid reservoir

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location C12N 5/04

Claims (4)

[Claims] 1. Proliferation of the biological cells by raising the culture solution below the tray in contact with the biological cells inoculated on the tray in the culture tank by raising them in a foam shape. Method for culturing biological cells. 2. The growth of a biological cell is attempted by repeating a cycle of temporarily suspending the foamy culture solution in contact with the biological cell and then again bringing the biological culture cell into contact with the foamy culture solution. The method described in. 3. A culturing apparatus having, in a culturing tank, one to a plurality of trays on which biological cells are placed, and a foaming device for foaming a culture solution contained below the trays. 4. A culture device comprising a culture tank and a foam-forming tank, wherein the culture tank has one to a plurality of trays on which biological cells are placed, and the foam-forming tank foams a culture solution. Have a foamer,
Between the culture tank and the foam forming tank, there is a foam sending pipe for sending the foam culture liquid to the culture tank, and a liquid sending pipe for sending the culture liquid accumulated in the lower part of the culture tank to the foam forming tank. Incubator.