JPH0367578A - Culture device - Google Patents

Abstract

PURPOSE:To obtain the title device of specific constitution preventing culture cells from growing into a large lump state by proper shear force resulting from turbulent flow and removing foams attached to cells by producing turbulent flow in flow of culture solution by aerated stirring. CONSTITUTION:First, air humidified into a saturated state by a humidifier 12 is sent from an intake vent 11a equipped at a bottom wall of a bottomed cylindrical culture container 1 through a porous material 4 fixed at the bottom of the container 1 to a culture solution 3 in a state of small foams. Successively, the small foams are raised together with the culture solution 3 in an air lift pipe 5 so that the culture solution 3 is recycled. Successively, the foams raised in the air lift pipe 5 are released through an exhaust column 6 to a gas phase and further from an exhaust vent 9 to the outside of the culture container 1 so that turbulent flow is produced in the flow of the culture solution 3 by limiting liquid level to part set in the exhaust vent 9 to constitute the objective device.

Description

DETAILED DESCRIPTION OF THE INVENTION [Object of the Invention] (Industrial Field of 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 for dedifferentiating plant cells and inducing callus, for example, an agar medium containing nutrients and hormones is placed in a Petri dish, plant cells are placed on the agar medium, There are methods to induce callus through dedifferentiation and proliferation, but
In this case, in order to efficiently dedifferentiate and proliferate plant cells, it is necessary to frequently replace the agar medium containing nutrients.

There is also a liquid culture method in which the callus is transferred to a culture medium in a culture container and cultured at the point when the cells of the plant dedifferentiate on an agar medium and become callus.
The agar medium was replaced every ~3 weeks, and even after being transferred to the culture medium, the culture medium was shaken to increase the concentration of molten oxygen in the culture medium.

It is necessary to aerate the culture solution by stirring.

Furthermore, there is a method of direct ventilation into the culture solution, but in this method, a vent hole is provided near the bottom wall of the container, air is sent into the culture solution from the outside through the vent hole, and the introduced air is small. The bubbles expand the contact area between the culture solution and air, thereby increasing the dissolved oxygen concentration in the culture solution.

(Problems to be Solved by the Invention) When aeration is carried out in the form of small bubbles, the small bubbles adhere to the plant cells being cultured, causing the plant cells to float near the water surface, making it impossible to uniformly stir the cells. A problem arises in that the attached air bubbles attract other cells, making the population of cells larger and larger.

The present invention provides a culture device that can apply shock to cells and air bubbles in order to remove air bubbles attached to cells in aeration using small air bubbles, and that can suppress cells from growing into large clumps. The purpose is to provide.

[Structure of the invention]

(Means for Solving the Problems) The present invention provides an air lift tube inside a culture container in which cells are cultured and proliferated using a liquid, and the culture container is provided with an air supply port, an exhaust port, a liquid supply port, a liquid drain port, etc. This culture device is characterized in that an exhaust pipe is provided above the air lift tube.

That is, in the present invention, in order to culture plant cells by aerating and stirring the liquid, the culture container has an air supply port near the bottom wall, a porous body is arranged to allow ventilation in the form of small bubbles, and , the culture container has an exhaust port.

A liquid supply port, a liquid drain port, etc. are provided, and an air lift tube is provided inside the container to facilitate the flow of the culture medium through aeration and agitation, and to restrict the flow of air to the upper part of the air lift tube. This is a culture device equipped with an exhaust pipe.

(Function) The air supplied from the air supply port passes through the porous body and is supplied into the culture solution in the form of small bubbles, and these small bubbles rise in the culture solution in the air lift tube, causing the small bubbles to rise. As it rises, the culture solution also rises, and the culture solution circulates by rising inside the air lift tube and descending outside the air lift tube.The air bubbles that rose inside the air lift tube further rise inside the exhaust stack, and air bubbles are removed from the liquid surface. It is released into the phase and out of the culture container from the exhaust port, and a liquid level is created at the part where the exhaust pipe is provided, preventing the rise of air bubbles and hindering the circulation of the culture solution.

(Example) Next, an example of the present invention will be described. Figure 1 shows the inside of a culture container formed into a cylindrical shape with a bottom, the culture solution 3 inserted into the culture container 1, and the porous body 4 provided at the bottom of the culture container 1.
, an air lift pipe 5 attached to the upper part of the porous body 4 in the vertical direction, and an exhaust pipe 6 attached to the upper part of the air lift pipe 5.

A lid 7 for fixing the exhaust pipe 6 to the culture container l, a port 8 for planting the lid 7, an exhaust port 9 opened for the lid 7, a filter 10 connected to the exhaust port 9, 1 shows a culture device consisting of an air supply port 11a provided on the bottom wall of a container 1 and a humidifier 12 connected to the air supply port 11a.

That is, 1 is a culture container formed into a cylindrical shape with a bottom made of glass or the like. This culture container 1 is filled with a culture solution 3 for growing plant cells 2, and a porous material is placed at the bottom of the culture container 1. 4, an air lift tube 5 is attached vertically to the top of the porous body 4, an exhaust tube 6 is supported from the lid 7 of the culture container 1 to the top of the air lift tube 5, and is provided with a planting port 8 for introducing plant cells and an exhaust port 9, and the exhaust port 9 is equipped with a sterilization filter 10 to prevent invasion of various germs.
a is connected, and an air supply port 11a is connected to the bottom wall of the culture container 1.
The other end of the air supply port 11a is connected to a humidifier 12, the humidifier 12 is filled with sterile water 13 for humidification, and the air supply port 11b is opened to the sterile water 13 of the humidifier 12. Air supply port 11
A sterilizing filter 10b and an air pump 14 are connected to b, and a liquid supply port 15 is provided on the upper wall of the culture container 1, and the other end of the liquid supply port 15 is connected to a culture solution supply tank 17. is opened into the culture solution 3 in the tank 17, and the culture solution 3
is fed into the culture container 1 using a roller pump 16, a vent 18 is provided in the culture solution supply tank 17, and a sterilization filter 10c is connected to the 1 vent 18. A drain port 19 for discharging the culture solution 3 is attached, and a valve 20 is connected to the drain port 19.

In this device, when growing plant cells, the pump 16 is first driven, and the culture solution 3 in the tank 17 is supplied to the culture container 1 through the solution supply port 15, so that the liquid level of the culture solution 3 in the culture container 1 is Adjust the liquid level so that it is inside the exhaust pipe 6,
Inject the plant cells to be propagated into the culture solution 3 from the planting port 8,
The air pump 14 is driven to supply air to the lower surface of the porous body 4 of the culture container 1 via the filter 10b and the humidifier 12.

Note that the air supplied to the lower surface of the porous body 4 is filtered through the filter 1.
It is sterilized and sterile in 0b, and humidifier 1
2, the humidity is almost saturated.

The air supplied to the lower surface of the porous body 4 by the air pump 14 passes through minute gaps in the porous body 4 and exits to the upper surface of the porous body 4, becoming minute bubbles and forming the culture solution 3 in the air lift tube 5.
The bubbles rise toward the liquid surface, accompanied by the culture solution 3, and near the liquid surface, all the bubbles gather inside the exhaust pipe 6. Since the space between the outside of the exhaust pipe 6 and the culture container 1 is closed, the air bubbles Therefore, the air is filled up to the height of the lower surface of the exhaust pipe 6, and the liquid level is kept constant, and this liquid level obstructs the upward flow of the culture solution 3 in the air lift tube 5, causing turbulence. The culture solution 3 descends on the outside of the airlift 5, causing a flow, the culture solution 3 in the culture container 1 is evenly stirred, and all air bubbles are collected in the exhaust pipe 6 and discharged to the outside of the culture container 1 through the filter 10a. .

By providing the exhaust pipe 6 in the culture container l, it is possible to limit the liquid level from which air bubbles in the liquid are discharged into the gas phase, and this limitation of the liquid level does not cause turbulence in the flow of the culture liquid 3. This turbulent flow applies an appropriate shearing force to the plant cells being cultured, releases air bubbles attached to the cells, and suppresses the development of large, clumpy growth of plant cells.

〔Effect of the invention〕

By installing an exhaust stack, turbulent flow is created.

Appropriate shearing force can be applied to the cells being cultured, thereby suppressing the phenomenon in which the cells grow in large clumps, and also making it possible to release air bubbles attached to the cells due to turbulent flow.

[Brief explanation of drawings]

FIG. 1 is a configuration diagram of a culture apparatus showing an embodiment of the present invention. 1...Culture container 3...Culture solution 4...
Porous body 5... Air lift pipe 6... Exhaust pipe 7... Lid 8... Planting port 12... Humidifier 9... Exhaust port

Claims (1)

[Claims] A container formed into a cylindrical shape with a bottom, a culture solution inserted into the container, a porous body provided at the bottom of the container, and an air lift tube attached vertically to the top of the porous body. , an exhaust pipe attached to the top of the air lift pipe, a lid that fixes the exhaust pipe to the container, a planting port opened in the lid, and an exhaust port opened in the lid;
A culture device comprising a filtration device connected to the exhaust port, an air supply port provided on the bottom wall of the container, and a humidifier connected to the air supply port.