JPH03160982A - Culturing device - Google Patents

Abstract

PURPOSE:To enhance efficiency of stirring and discharging of callus by providing depressed part in bottom part performing introduction of air and discharging of callus, etc., and inserting porous body having specific structure in vertically movable state in a culturing device installing cylindrical air lift tube in culturing vessel and performing stirring with ventilation. CONSTITUTION:Culturing solution 4 and plant cell are charged in a culturing vessel 15 and air is supplied to bottom part of depressed part 16 formed in bottom part of the vessel 15 from an air supplying tube 19, then air bubble is generated from the bottom part of the vessel 15, thus the plant cell is cultured with circulating the culturing solution 4 in the action of an air lift tube 21 with giving oxygen to the culturing solution 4. On the other hand, the culturing solution 4 is discharged from a discharging pipe 17 connected to bottom part of the depressed part 16. Furthermore, a porous body 20 having holes 29 and 30, etc., having fixed sizes in every points is inserted in the depressed part 16 in vertically movable state. By said method, ventilation can be performed from the lowest part of the culturing vessel 15 and discharging of culturing solution 4 only or discharging of culturing solution 4 containing callus can be performed.

Description

DETAILED DESCRIPTION OF THE INVENTION [Objects of the Invention] 1 (Field of Industrial Application) The present invention relates to a culture device for multiplying plant cells in large quantities using a culture solution.

(Prior art) As a general method to dedifferentiate plant cells and induce callus, for example, an agar medium containing nutrients and hormones is placed inside a Petri book, and plant cells are placed on the agar medium to dedifferentiate. , a method of proliferating and inducing callus is known.

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

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

In this case, for example, even after changing the agar medium every 2 to 3 weeks and transferring it to the culture medium, the culture medium may be shaken or stirred to increase the concentration of molten oxygen in the culture medium. ventilation is being carried out.

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

In this case, a porous hole is provided near the bottom wall of the culture container, and the culture medium is aerated through this porous member to generate small air bubbles in the culture medium, which are then raised to the water surface. This movement 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.

(Problem to be solved by the invention) However, in the conventional direct ventilation method described above,
Since the callus is heavier than the culture solution, the callus sinks into the culture solution.

Therefore, in order to efficiently cause callus to circulate in the culture solution, air bubbles must be generated from the lowest part of the culture solution.

Furthermore, when taking out callus from a culture container, it is necessary to take it out from the lowest part of the culture container where the callus is submerged, otherwise it will not be possible to remove all of the callus from the culture container without leaving any callus behind.

In other words, unless the two functions of aeration and discharge are carried out from the lowest part of the culture container, it is not possible to grow callus efficiently or to discharge it efficiently.

For this reason, there has been a strong desire to develop a culture device with a bottom structure that does not allow callus to pass through when performing aeration or discharging only the culture solution, and that allows callus to be taken out when necessary.

In view of the above circumstances, the present invention provides a culture device that can ventilate from the bottom of the culture container and also discharge only the culture solution or the culture solution containing callus from the same part. It is intended to.

[Structure of the invention]

(Means for Solving the Problems) In order to achieve the above object, the culture device according to the present invention has the following features:
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 provide oxygen to the culture solution in the culture container. In a culture device that circulates water, a recess formed at the bottom of the culture container, an air supply pipe that supplies air to the bottom side of the recess, an exhaust pipe connected to the bottom of the recess, and a recess specified for each part. The present invention is characterized by comprising: a porous body having a recess of a large size formed therein and fitted into the recess so as to be vertically movable; and a position adjustment mechanism for adjusting the position of the porous body according to the state of use.

(Function) In the above configuration, by determining the position of the porous body using the position adjustment mechanism, the air supplied from the air supply pipe can be released from the porous body as microbubbles, or only the culture solution in the culture container can be released. The callus is discharged from the discharge pipe, or the callus is discharged together with the culture solution in the culture container from the discharge pipe.

(Example) FIG. 1 is a diagram showing the structure of an example 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. Bubbles are generated in the culture solution 4 in the device main body 1 to cause plant cells in the culture solution 4 to grow.

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 7 that humidifies the air that has been sterilized by the sterilization filter 6. The humidifier 8 is provided with a humidifier 8 that takes in air from the atmosphere, sterilizes it, humidifies it, and then supplies it to the culture device main body 1.

The culture solution supply device 3 includes a culture solution supply tank 10 filled with culture solution 9, a ventilation pipe 12 that communicates the culture solution supply tank 10 with the atmosphere via a sterilization filter 11, and the culture solution supply tank 10. The roller pump 13 takes in the culture solution 9 in the culture solution supply tank 10 and supplies it to the culture device main body 1, and when a culture solution supply command is supplied, the roller pump 13 pumps the culture solution 9 in the culture solution supply tank 10. The liquid 9 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, a recess 16 formed at the bottom of the culture container 15, and a discharge pipe 17 connected to the bottom of the recess 6. , a valve 18 attached to this discharge pipe 17 , one air supply pipe connected to the side surface of the recess 16 , a porous body 20 fitted into the recess 16 so as to be vertically movable, and this porous body 20 A cylindrical air lift pipe 21 is provided above the air lift pipe 21 so that its axis coincides with the vertical direction.

Furthermore, the culture device main body 1 includes a plate-shaped lid 23 that closes the top of the culture container 15, a planting port 24 provided in the lid 23, and a sterilization filter 25 that allows the top of the culture container 15 to be exposed to the atmosphere. a ventilation pipe 26 communicating with the lid 23;
is attached to the porous body 2 so as to be movable up and down.
0, and a support rod 27 connected to 0.

In this case, as shown in FIG. 2, the porous body 20 has a large number of micropores 28 for generating microbubbles in its upper part, and a plurality of small holes 29 in its upper side that allow only the culture solution 4 to pass through. is formed, and a plurality of holes 30 are formed on the lower side of the side for allowing the callus and the culture medium 4 to pass through.

The culture device main body 1 is replenished with the culture solution as appropriate by the culture solution supply device 3, and air bubbles are generated in the culture solution 4 in the culture device main body 1 by the air supplied from the air supply device 2, thereby culturing. Grow plant cells in Solution 4.

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

When growing plant cells using this culture device, first of all, inside the culture container 15, each sterilization filter 6, 11,
After the parts 25 and the like are sterilized and made aseptic, the support rod 27 is lowered and the entire porous body 20 is fitted into the recess 16 as shown in FIG. 3(a).

Next, the roller pump 13 is driven to take in the culture solution 9 in the culture wave supply tank 10 and continuously supply the culture solution 4 into the culture container 5 until the liquid level of the culture solution 4 reaches a predetermined height.

Next, after the plant cells to be propagated are introduced from the planting port 24 and placed in the culture solution 4, the air bomb 5 is driven to take in outside air, which is sterilized and humidified. It is supplied to the recess 16 of the culture container 1-5.

As a result, small bubbles 31 come out from the micropores 28 of the porous body 20 and rise through the inside of the air lift tube 21, and the movement of the bubbles 31 causes the culture solution 4 on the bottom side of the culture container 15 to After being moved upward and flowing in the outer circumferential direction near the water surface, it passes through the outside of the air lift tube 21 and returns to the bottom side of the culture container 15.

Thereafter, while air is being supplied from the air supply device 2, the above-mentioned circulation operation of the culture solution 4 is repeated, and the culture solution is stirred, oxygen is given to the culture solution 4, and the culture solution 4 is Cells inside can grow efficiently.

In addition, when draining 4 volumes of the culture solution in the culture container 15, the air supply operation of the air supply device 2 is stopped, and after the valve 18 is opened, the support rod 27 is slightly lifted. As shown in FIG. 3(b), a porous body 20
The upper side of is made to protrude upward from the recessed part 16.

As a result, the culture solution 4 in the culture container 15 flows into the recess 16 through each small hole 29 of the porous body 20, and is discharged via the discharge pipe 1-7.

In this case, each small hole 29 is smaller than the plant cell, so the plant cell is blocked by each small hole 29 and does not flow into the recess 16.

When each small hole 29 becomes clogged with plant cells, the air supply operation of the air supply device 2 is restarted to perform backwashing with air.

Thereafter, when the draining of the culture solution 4 is completed, the support rod 27 is lowered and the porous body 20 is moved into the recess 1 as shown in FIG. 3(a).
6 is inserted again.

In addition, when discharging callus together with the culture solution 4 in the culture container 15, the air supply operation of the air supply device 2 is stopped, and after the valve 18 is opened, the support rod 27 is completely lifted up. Figure 3 (c
), the lower side of the porous body 2o is made to protrude upward from the recess 16.

As a result, the callus flows into the recess 16 together with the culture solution 4 in the culture container 15 through each hole 3o of the porous body 2o, and is discharged via the discharge pipe 17.

As described above, in this embodiment, the micropores 28 are formed in the upper part of the side, and the passage of the callus is blocked on the side upper side to prevent the passage of the culture solution 4.
A plurality of small holes 29 are formed to allow only the callus to pass through, and a plurality of holes 3o are formed at the lower side of the side to allow the callus and the culture solution 4 to pass through.
The porous body 20 formed with the above structure is fitted into the recess l6 formed at the bottom of the culture container 15 so as to be able to move up and down.
It is possible to ventilate the culture solution 4 from the bottom of the culture container 15 to thoroughly stir the culture solution 4 to every corner, and also to drain only the culture solution 4 completely or to drain the culture solution 4 containing callus from the same part. Can be done.

Furthermore, even if the porous body 2o becomes clogged, the clog can be cleared by backwashing through ventilation.

〔Effect of the invention〕

As explained above, according to the present invention, ventilation can be carried out from the lowest part of the culture container, and only the culture solution or the culture solution containing callus can be discharged from the same part.

[Brief explanation of the drawing]

FIG. 1 is a schematic diagram of an embodiment of the culture device according to the present invention, FIG. 2 is a detailed perspective view of the porous body shown in FIG. 1,
FIG. 3 is a sectional view for explaining the operation of the culture apparatus according to the present invention. 4...Culture solution 15...Culture container 16...Recess 17...Discharge pipe 19...Air supply pipe 21...Air lift tube 20...Porous body 27...Position adjustment Mechanism (support rod)

Claims (1)

[Claims] (1) A cylindrical air lift tube is provided inside the 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 give oxygen to the culture solution in the culture container. This culture device that circulates the culture solution includes a recess formed at the bottom of the culture container, an air supply pipe that supplies air to the bottom of the recess, an exhaust pipe connected to the bottom of the recess, and each part. A porous body having a hole of a specified size and fitted into the recess so as to be vertically movable; and a position adjustment mechanism that adjusts the position of the porous body according to usage conditions. A culture device that uses