JPH02265468A - Rotating bottle type culture device - Google Patents

Abstract

PURPOSE:To directly detect a culture medium in rotating bottle by detectors and make it possible to control the culture medium by providing a detector and nozzle for passing through a lid part of the rotating bottle and directly detecting and controlling conditions of the culture medium in the bottle. CONSTITUTION:A ventilation nozzle 4 having a plurality of pores is arranged in a rotating bottle 1 and the nozzle 4 is communicated and connected to an air pump 5 and air is fed in a form of fine foam from the ventilation nozzle 4 into a culture medium. A nozzle 6 arranged into the rotating bottle 1 is provided in a discharge port 7 having a filter and the interior of the bottle 1 is kept to ordinary pressure by opening the interior to the air. A nozzle 9 is further arranged in the rotating bottle 1 and supply, exchange, etc., of the culture medium is carried out by the nozzle 9. On the other hand, a detecting rod in which a hydrogen ion concentration detector 11 and dissolved oxygen concentration detector 13 are integrated is inserted into the rotating bottle 1 and the detecting rod is connected to a controller 12 and controller 14.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a rotary bottle culture device, and particularly to a rotary bottle culture device filled with a liquid medium and used to culture tissues and cells of animals and plants.

[Prior Art] A rotary bottle culture device called a roller bottle type, which is filled with a liquid medium and cultured animal and plant tissues, cells, etc. in a rotating state, has been known.

To explain this conventional rotary bottle type culture device with reference to FIG. 3, the inner diameter of the bottle mouth is significantly smaller than the inner diameter of the body, and the rotary bottle 1 is made of glass and is formed by blow-blasting. 2
The bottle opening 1a is blocked by the bottle opening 1a. Reference numeral 3 indicates the medium liquid level, which is located slightly below the bottle opening 1a.

A ventilation nozzle 4 having a plurality of pores is disposed inside the rotary bottle l, and this ventilation nozzle 4 is connected to an air pump 5. When the air pump 5 operates, air is passed through the ventilation nozzle 4 into fine particles. It is supplied into the culture medium as foam.

Note that by supplying air from the ventilation nozzle 4, the rotary bottle I
If the air pressure inside the bottle becomes high, the air inside the bottle will flow through the nozzle 6.
The air is discharged through the exhaust lower, which is equipped with a filter to prevent germs from entering from the outside.

Reference numeral 8 denotes a separation tank, and a rotary plate 1 is installed at the bottom of the separation tank 8.
A suction nozzle 9 extending into the interior is provided.
Further, a supply nozzle 6 having a roller pump 10 is provided at the upper part of the separation tank 8, and this supply nozzle 6 extends into the rotary plate 1 as described above.
Then, as the roller pump 10 operates, the supernatant liquid in the separation tank 8 is sucked and sent from the supply nozzle 6 into the rotary tray 1, and the pressure in the separation tank 8 decreases accordingly. The culture medium in the tank 1 is sucked through the suction nozzle 9 and replenished into the separation tank 8.

The roller pump 10 operates intermittently, and when the operation is interrupted, exhaust is performed from the exhaust lower.

The separation tank 8 has the function of separating the culture product and the medium solution, precipitating the product, and turning the culture solution into a supernatant when the culture is completed.

In addition, a hydrogen ion concentration detector (PH sensor) 11 is disposed in the separation tank 8, and the detector 11 detects the hydrogen ion concentration of the culture medium in the separation tank 8.
2, a hydrogen ion concentration adjusting solution is supplied into the separation tank 8 as necessary for adjustment.

Inside the separation tank 8, there is also a dissolved oxygen concentration detector (DO sensor)! 3 is arranged, the detector 13 detects the dissolved oxygen concentration of the culture medium in the separation tank B, and the controller 14 adjusts the amount of air supplied by the air pump 5 as necessary.

[Problems to be Solved by the Invention] However, in the conventional apparatus described above, only a simple sealing material is interposed between the bottle mouth and the non-rotating part, and little attention is paid to water leakage. This is because the culture solution was only filled to the bottom of the relatively small bottle mouth.

Accordingly, it has been impossible to increase the size of the non-rotating portion provided at the bottle mouth.

Therefore, as shown in Fig. 3, a separation tank 8 is provided, and the hydrogen ion concentration and dissolved oxygen concentration are detected in the separation tank 8. There is a problem that there is a delay and the adjustment fluctuation range of the culture solution in the one-rotation dish 1 has to be large.

Further, in order to correlate the condition in the separation tank 8 with the condition in the rotary plate 1 as much as possible, the roller pump IO must be operated intermittently.

This requires frequent interruption of the exhaust from the exhaust lower and circulation of the culture solution containing tissues or cells in the process of culturing from the suction nozzle 9 through the separation tank 8, the roller pump 10, and the supply nozzle 6 back to the rotating plate l. There is a problem in that this is not preferable for culture because it requires frequent steps and causes unnecessary fluctuations in the tissues and cells that are being cultured.

The present invention was devised in view of the above-mentioned circumstances, and its purpose is to provide a rotating dish culture device that can directly detect and adjust the culture solution in a rotating bottle using detectors. There is a particular thing.

[Means for Solving the Problems] In order to achieve the above-mentioned object, the present invention comprises storing an object to be cultured, such as an animal or plant tissue or cell, and a culture solution in a rotating bottle,
In the rotating dish type culture device that promotes culture while rotating the rotating dish, the inner diameter of the bottle mouth of the rotating dish is configured to be below the surface of the culture liquid, and the lid body that closes the bottle mouth is made of a sealing material. A detector and nozzles are attached to the rotating bottle in a non-rotating state regardless of the rotation of the rotating plate through the lid, and penetrate the lid to directly detect and adjust the condition of the culture solution in the bottle. It is characterized by the fact that it has been provided.

"Function" According to the present invention, the state of the culture solution in the one-turn bottle can be directly detected by the above-mentioned means using detectors.

There is no time delay in the detected values, and the culture solution can be quickly adjusted, and there is no need to circulate the culture solution to the outside, allowing stable culture.

[Example] Hereinafter, the rotary dish culture device according to the present invention will be described as shown in FIGS. 1 and 2.
This will be explained with reference to the figures.

FIG. 1 shows a basic configuration diagram of a rotating dish type culture device, and a ventilation nozzle 4 having a plurality of pores is arranged in one rotating dish l.
This ventilation nozzle 4 is connected to an air pump 5, and when the air pump 5 is operated, air is supplied from the ventilation nozzle 4 into the culture solution in the form of fine bubbles.

A nozzle 6 disposed inside the rotary bottle 1 is connected to an exhaust lower with a filter, and the inside of the bottle 1 is opened to the atmosphere and maintained at normal pressure.

Further, a nozzle 9 is disposed inside the 5-turn bottle l, and the nozzle 9 is used to replenish, replace, etc. the culture solution.

- On the other hand, a detection rod that integrates a hydrogen ion concentration detector 11 and a dissolved oxygen concentration detector 13 is inserted into the rotary bottle l, and the detection rod is connected to the controller 12 and the controller 14.

FIG. 2 shows a sectional view of the rotary bottle 1 and each device attached to the rotary bottle 1.

A rotary lid 15 is integrally fixed to the bottle spout 1a of the rotary bottle 1, which is cylindrical with a bottom and whose inner diameter is equal to the inner diameter of the bottle body, by a fixing band 17 via a backing 16.

The rotary lid 15 is made of an annular member, and has a central opening 15a.
An inner fixed lid 18 is fitted on the inside.

A backing 19 is provided between the rotary lid 15 and the inner fixed lid 18.
is interposed so that the rotary lid 15 rotates together with the rotary bottle l, while the inner fixed lid 18 remains stationary.

Furthermore, sealing materials 2o and 2.degree. are interposed on the inner and outer surfaces of the rotating lid 15 to prevent liquid leakage from the contact surface between the one-turning lid 15 and the inner fixed lid 18.

On the other hand, an outer fixed lid 21 is integrally fixed to the outer surface of the inner fixed lid 18 with screws 22.
A rotation stopper 24 protruding from the fixing base 23 is fitted into I, and is fixed by a set screw 25.

The inner and outer fixing members 24 are fixed in a stationary state regardless of the rotating bottle l and the rotating lid 15.

Then, the nozzle 4. is inserted through the inner and outer fixed lids 18.21.
6.9 is installed.

In addition, a hydrogen ion concentration detector II is installed on the outer fixed lid 2 irises.
A holding plug 27 holding a detection rod incorporating a dissolved oxygen concentration detector I3 is attached.

In the configuration described above, the separation tank 8 and roller pump IO from the conventional apparatus shown in FIG. 3 are no longer needed and are removed.

The culture work is carried out by rotating the rotary bottle 1 by driving the rollers 26 on the fixed table 23 without circulating the culture solution. is detected.

Note that glass or transparent plastic can be used for the one-turn bottle, and metal, hard synthetic resin, etc. can be used for the lid.

Materials for sealing and backing materials include nitrile rubber, silicone rubber, pyton, chloroprene rubber, polyacrylic rubber, bibalon, skillen rubber, butyl rubber,
Teflon or the like can be used.

In addition, in the one-piece embodiment, two types of detectors were described in which the culture top is also important, but auxiliary detectors such as a temperature detector can be further installed.

The non-rotating portion of the lid should have a minimum diameter that is necessary and sufficient for installing the detector and nozzles.

[Effects of the Invention] As is clear from the description of the embodiments, according to the present invention, the inner diameter of the bottle opening of the rotary bottle is configured to be below the surface of the culture solution, and the lid body that closes the bottle opening is configured. Since a direct detector etc. is installed in the non-rotating part, the condition of the culture solution in the rotating bottle can be directly detected by various detectors, there is no time delay in the detected value, and the culture solution can be adjusted quickly. .

In addition, according to the present invention, there is no need to circulate the culture solution between the rotating bottle and the separation tank, so that unnecessary fluctuations are not caused to tissues and cells in the process of culturing that occur due to circulation, and stable culture can be performed. It will be done.

Moreover, by removing the separation tank, roller pump, etc., the apparatus can be simplified and the operation can be simplified, and the possibility of contamination of the culture solution with germs is reduced.

[Brief explanation of drawings]

FIG. 1 is a basic configuration diagram showing an embodiment of a rotary bottle type culture device according to the present invention, FIG. 2 is a sectional view of the rotary bottle of the same device, and FIG.
The figure is a basic configuration diagram of a conventional rotary bottle type culture device. 4. 11 ・ 12゜ 13 ・ 15 ・ 18 ・ 21 ・ 27

Claims (1)

[Claims] (1) In a rotary bottle type culture device in which an object to be cultured, such as an animal or plant tissue or cell, and a culture solution are stored in a rotary bottle and the culture is promoted while rotating the rotary bottle, the inner diameter of the bottle mouth of the rotary bottle is is configured to a size that is below the surface of the culture solution, and a lid that closes the bottle opening is attached to the rotating bottle through a sealing material in a non-rotating state regardless of the rotation of the rotating bottle, and the lid part 1. A rotary bottle-type culture device characterized by being provided with a detector and nozzles for directly detecting and adjusting the state of the culture solution in the bottle by penetrating the bottle.