JPH04258285A - Culture device - Google Patents

Abstract

PURPOSE:To prevent plant cells from largely growing into a lump state by applying proper shear force to plant cells in a culture solution while aerating with small bubbles and stirring. CONSTITUTION:In a culture device wherein a cylindrical air lift pipe 18 is laid in the interior of a culture container 15 of culturing and multiplying cells by using a culture solution 4 and the culture solution 4 is circulated while adding oxygen to the culture solution 4 existing in the culture container 15 of generating bubbles 27 from the bottom of the culture container 15, a water flow stirring part (rotary blade 19) is laid in the air lift pipe 18 and flow of the circulating culture solution 4 is made weakly turbulent by the water flow stirring part.

Description

[Detailed description of the invention]

[Object of the invention]

0002

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culturing apparatus for multiplying plant cells in large quantities using a culture solution.

0003

[Prior Art] As a general method for dedifferentiating plant cells and inducing callus, for example, an agar medium containing nutrients and hormones is placed in a Petri dish, and plant cells are placed on the agar medium and dedifferentiated. A method of inducing callus by differentiation and proliferation is known.

[0004] In this case, in order to efficiently dedifferentiate and proliferate plant cells, the agar medium containing nutrients is frequently replaced.

[0005] Furthermore, a liquid culture method is also known in which, once the cells of a plant are dedifferentiated on an agar medium and become callus, the callus is transferred to a culture solution in a culture container and cultured.

[0006] In this case, for example, the agar medium is replaced every 2 to 3 weeks, and even after it is transferred to the culture medium, it is shaken or stirred to increase the concentration of molten oxygen in the culture medium. The culture solution is aerated.

[0007] A method of increasing dissolved oxygen by directly aerating the culture solution is also known.

[0008] In this case, a porous member is provided near the bottom wall of the culture container, and the culture medium is aerated through the porous member to generate small air bubbles in the culture liquid, which are then raised to the water surface. The movement of the bubbles generates a flow in the culture solution to agitate the culture solution, and also increases the amount of dissolved oxygen in the culture solution to efficiently provide oxygen and nutrients to the cells in the culture solution.

[0009]

[Problems to be Solved by the Invention] However, in the above-mentioned conventional direct aeration method, small air bubbles are used in order to increase the contact area between the air and the culture solution and to generate a flow in the culture solution. However, since the shearing force of such small bubbles is small, the group of plant cells becomes large and large, and even if they are removed, it is difficult to redifferentiate into the original plant, and one root may have only 2 to 3 buds. In some cases, the plants had two to three roots and one bud, and the number of roots and buds were not the same.

[0010] In view of the above circumstances, the present invention aims to apply an appropriate shearing force to plant cells in a culture solution while performing aeration and agitation using small air bubbles, thereby suppressing the plant cells from growing in large clumps. The purpose is to provide a culture device that can.

[Configuration of the invention]

0012

[Means for Solving the Problems] In order to achieve the above object, a culture device according to the present invention is provided with a cylindrical air lift tube inside a culture container for culturing and proliferating cells using a culture solution, and In a culture device that circulates the culture solution while giving oxygen to the culture solution in the culture vessel by generating air bubbles from the bottom of the culture vessel, a water flow stirring section is provided in the air lift tube, and the water flow stirring section is used to circulate the culture solution. The feature is that the flow of the culture solution is disturbed.

[0013]

[Operation] In the above structure, while the culture solution is moved by air bubbles from the bottom of the culture container, shear force is generated by the water agitation section to suppress clumping of plant cells in the culture solution.

[0014]

Embodiment FIG. 1 is a block diagram showing an embodiment of a culture apparatus according to the present invention.

The culture device shown in this figure is equipped with a culture device main body 1, an air supply device 2, and a culture solution supply device 3. 2, the culture solution 4 in the culture device main body 1
Air bubbles 16 are generated therein to grow the plant cells in the culture solution 4.

The air supply device 2 includes an air pump 5 that takes in air, a sterilization filter 6 that removes germs from the air taken in by the air pump 5, and a sterile water that converts the air sterilized by the sterilization filter 6 into sterile water. 7 and a humidifier 8 for supplying the humidified air to the culture apparatus main body 1, which takes in air from the atmosphere, sterilizes it, humidifies it, and then supplies it to the culture apparatus main body 1.

The culture solution supply device 3 includes a culture solution supply tank 10 filled with the culture solution 4, a ventilation pipe 12 that communicates the culture solution supply tank 10 with the atmosphere via a sterilization filter 11, and a culture solution supply tank 10 filled with the culture solution 4. Culture solution 4 in the solution supply tank 10
and a roller pump 13 that takes in and supplies it to the culture device main body 1, and when a culture solution supply command is supplied, the roller pump 13 supplies the culture solution to the culture solution supply tank 1.
0 is taken in and supplied to the culture apparatus main body 1.

The culture device main body 1 includes a bottomed cylindrical culture container 15 made of glass or the like, and a valve 20 provided on the bottom wall of the culture container 15 to open the culture container 1.
Discharge pipe 21 for discharging culture solution 4, callus, etc. in 5
a recess 16 formed at the bottom of the culture container 15 to take in the air supplied from the air supply device 2; The porous body 17 generates small bubbles 27 in the liquid 4.

Furthermore, this culture device main body 1 includes a cylindrical air lift tube 18 disposed above the porous body 17 so that its axis coincides with the vertical direction, and a rotating tube inside the air lift tube 18. A rotary blade 19 that is freely provided, a plate-shaped lid 23 that closes the top of the culture container 15, and this lid 23.
and a ventilation pipe 26 that communicates the upper part of the culture container 15 with the atmosphere through a sterilization filter 25.

[0020] Then, as appropriate, the culture solution supply device 3
While the culture solution 4 is replenished, air bubbles 27 are generated in the culture solution 4 in the culture device main body 1 by the air supplied from the air supply device 2, and the rotary blade 19 is rotated while circulating the culture solution 4. The plant cells in the culture solution 4 are allowed to grow by suppressing the plant cells from growing in large lumps.

Next, the operation of this embodiment will be explained with reference to FIG.

When growing plant cells using this culture device, first of all, the roller pump 13 is driven to take in the culture solution 4 in the culture solution supply tank 10, and
This is supplied into the culture container 15 to raise the level of the culture solution 4 to a predetermined height.

Next, plant cells to be propagated are introduced through the planting port 24 and then introduced into the culture medium 4 through the opening 14.
After the culture container 15 is placed inside, the air pump 5 is driven to draw in outside air, and the air is sterilized, humidified, and supplied to the recess 16 of the culture container 15.

As a result, small air bubbles 27 come out from the porous body 17 and rise through the inside of the air lift tube 18, and the movement of the air bubbles 27 causes the culture solution 4 on the bottom side of the culture container 15 to move upward. The rotary blades 19 are rotated while moving, and the shear force generated at that time suppresses the plant cells from growing in large chunks. When the culture solution 4 that has moved upward reaches the water surface, it passes through the outside of the air lift tube 18 and is returned to the bottom side of the culture container 15.

Thereafter, while air is being supplied from the air supply device 2, the above-described circulation operation of the culture solution 4 is repeated, and the culture solution 4 is stirred and oxygen is given to the culture solution 4. The cells in the culture medium 4 grow efficiently.

When the plant cells in the culture solution 4 reach a desired growth level, the air supply operation of the air supply device 2 is stopped, and then the valve 20 is opened and the culture solution 4 is discharged through the discharge pipe 21. At the same time, plant cells are excreted.

In this way, in this embodiment, the bubbles 2
The rotating blades 19 are rotated by the flow of the culture solution 4 being moved by the rise of the cell 7 to apply shear force to the plant cells. Appropriate shearing force can be applied to plant cells, thereby suppressing the plant cells from growing into large clumps.

Furthermore, in the above-described embodiment, since the rotating blades 19 are rotated by the rising force of the culture solution 4, the shearing force against small plant cells is reduced and the shearing force against large lumpy plant cells is reduced. can be increased, thereby allowing plant cells to proliferate efficiently.

[0029]

[Effects of the Invention] As explained above, according to the present invention, an appropriate shearing force is applied to the plant cells in the culture solution while performing aeration and agitation using small air bubbles, thereby causing the plant cells to grow in large clumps. can be suppressed.

[Brief explanation of the drawing]

FIG. 1 is a configuration diagram showing an embodiment of a culture device according to the present invention.

[Explanation of symbols]

4 Culture solution 14 Opening 15 Culture container 18 Air lift pipe 19 Water stirring part (rotating blade) 27 Air bubbles

Claims (1)

[Claims] 1. A cylindrical air lift tube is provided inside a culture container in which cells are cultured and proliferated using a culture solution, and air bubbles are generated from the bottom of the culture container to supply oxygen to the culture solution in the culture container. A culture device that circulates the culture solution while feeding the culture solution, characterized in that a water flow stirring section is provided in the air lift tube, and the water flow stirring section disturbs the flow of the culture solution during circulation.