SU1242518A1 - Device for growing cells or hybridoms - Google Patents

Description

124251

2. Installation according to claim 1, in connection with the fact that, in order to improve the cultivation of the hybrid, the installation is equipped with a pneumatic valve connecting the cultivator with the gas supply system to the remote chamber for contact liquid and gas, while the latter is connected by an intermediary. The invention relates to a technique of cell cultivation and can be used in cell biology, virology, medicine, and veterinary medicine.

The purpose of the invention is to more evenly distribute cells or hybridomas over the growth surface and improve mass transfer in the cell-body region, as well as increase the duration of hybridoma cultivation.

Fig. 1 is a schematic of a plant for the cultivation of hybridomas; . 2 is a diagram of a plant for culturing dependent cells; Fig. 3 shows a cultivator, an embodiment; 4 is a diagram of a remote chamber for contacting fluid and pelvis.

The installation includes a cultivator 1 with a nozzle 2 for introducing the culture fluid of cells, a nozzle 3 for introducing a gas mixture connected to a bacterial filter 4 of the gas mixture supply system, a nozzle 5 for evacuating the gas, a nozzle 6 for introducing the medium. The cultivator is provided with a hollow cup 7, inside of which there are baffles 8 and 9 along the height, which alternately have openings for fluid flow located in the central and peripheral parts.

The installation is also equipped with a remote chamber 10 for contacting liquid and gas, the upper part of which is connected to the upper part of the glass 7, and the lower part is connected to the cultivator 1, while in the upper chamber 10 there are plates 11 having openings with the shoulders 12 for the passage of gas.

In the case of cultivation of hybrids (Fig. I), the plant contains such a peristaltic pump to a vessel with a nutrient medium.

3. Installation under item 1, about tl and - cup, and so that when growing cells, the upper part of the glass is connected to the upper part of the external chamber for contact of liquid and gas by means of a peristaltic pump.

the same pneumatic valve 13 with the control unit 14. A pneumatic valve 13 connects the cultivator I to a gas supply system 15 and a remote chamber 10 for contacting liquid and gas. The remote chamber 10 is connected via the peristaltic pump 16 to the vessel 17 with the nutrient medium and through the valve 18 is connected to the cultivator 1. In addition, the lower part of the external chamber is connected to the cultivator 1 through the pneumatic valve 13 and through the locking device 19 to the sampling device 20,

The cultivator 1 is connected through pipe 21, pipe 2 and the peristaltic pump 22 to a collector 23 of the culture medium, which is connected to the atmosphere through a filter 24. The cultivator 1 through the pipe 3, the pneumatic valve I3, the bacterial filter 4 is connected to the gas supply system 15, which contains gas metering units 25 and 26, a gas flow regulator 27

mix and gauge 28.

In the case of culturing cells (Fig. 2) that are dependent on attachment, the upper part of the external chamber 10 for contacting liquid and gas is connected to the upper part of glass 7 by means of a peristaltic pump 16 and through a locking device 29 s: a bacterial filter 30. A cultivator 1 through a locking device 31

connected to the bacterial filter 4, and through the locking device 32 to the vessel 23 for the inoculum, which through the locking device 34 is connected to the bacterial filter 4, and through

locking device 35 with a bacterial filter 30.

Preferably, the design of the cultivator (fig.Z), where partitions 9

31

have the form of a cup with shoulders and holes in the central part, and the partitions 8 are made with a smaller diameter and form a gap with the walls of the glass 7. The partitions are fastened in pairs in the central part.

 The remote chamber 10 for contacting liquid and gas (Fig. 4) is made with extensions in the upper part, in which the plates 11 are located, having openings with shoulder 12 for the passage of gas

An apparatus for culturing attachment-dependent cells (Fig. 2) operates as follows.

Cultivator cavity 1. and hermetic cup 7 are filled with a gas mixture consisting of air and 5% COj before filling with medium. so that the pH of the medium is not changed during the filling. To do this, shut off the locking devices 32 and 34, the nozzle 2 and the gas through the filter 4 and the open locking device 31 enters through the nozzle. compressed air through the filter 4 and the open nozzle 2 with cells enters the cultivator 1 when the nozzles 3.5 and 6 are blocked and fills the space between the disks.

Then, the upper part of the sloping glass 7 is crossed and the culture is kept for 4-5 hours, which is necessary for cell attachment.

After this time, the peristaltic pump 16 is turned on with the openings 5 and 6 and the shut-off devices 32 and 34 closed. The medium circulates through the pump 16, the remote chamber 10, flows down into the cultivator 1 and moves upwards through the labyrinths of the cup 7. Circulation of the medium lasts as much time as is established for growing a specific type of cell culture.

At the end of the experiment, the culture medium is drained into the vessel 33 through the nozzle 2, while the locking device 29 is closed, and the locking device 32 and 35 are open and the gas phase flows through the bacterial filter. 4 under the pressure of 150-300 mm water column, and the liquid through the pipe 2 enters the vessel 33.

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The plant (Fig. 1) for the cultivation of hybridomas works as follows.

Before starting work, the cultivator 1 and chamber 10 are ventilated with a gas mixture (air and 4-6% COg) to exclude a change in the pH of the culture medium after placing it in the cultivator 1 from the vessel 33. The composition of the gas phase is selected, changing the capacity of the feeders 25 and 26. Valve 18 serves as a pneumatic resistance and provides pressure in the cultivator 1 by 2/3 more than in the chamber 10.

Then the cultivator 1 is filled with a nutrient medium with hybridomas, while the pump 16 is at work, it thaws, the locking device 32 is open and 31 is closed, while the gas mixture from filter 4 to nozzle 3 does not enter the cultivator 1 - blocked by the device 31. Gas mixture. enters through the filter 4 from the gas supply system 15, and pressure is created in the vessel 33, under the action of which the culture medium flows through the nozzle 2 into the cultivator 1, and thus the cup 7 is filled until the medium appears in its upper part. Then the locking device 32 is closed and the device is in such a state for the entire period of attachment of the hybridomas to the growth surface of the disks (30 hours).

After the hybridomas are attached, the pneumatic valve 13 is turned on, at that the nozzles 5 to 6 are closed and the nozzle 3, the liquid under the action of the gas phase pressure, opens through the upper part of the stack and 7 partially

is displaced into chamber 10, where a gas mixture is pressed into the chamber, filling the lower part of chamber 10. The pneumatic valve 13 is switched, the pipe 3 and the upper part of the hollow cup 7 are closed, and the liquid from chamber 10 flows through pipe 6 cultivator 1, displacing the gas phase through nozzle 5. Such cycles are repeated throughout the experiment. At the moment of filling the chamber 10, the device 19 is opened for a short time, during which the sample enters the sampler 20.

To ensure long-term cultivation, hybridomas are periodically or continuously replaced.

nutrient medium. By means of the pump 16, fresh medium is supplied, and by means of the pump 22, the culture is removed on the medium with the cells while maintaining the constant level of liquid in the cultivator. This is ensured by the fact that the pump 22 has a greater capacity than the pump 16, while the pipeline 21 is set at a predetermined level.

Thus, the proposed installation in comparison with the known ensures uniform planting of cells

or a hybrid on the growth surface and improves mass transfer in the cell-cell region. In the case of cultivation of cells dependent on attachment to the substrate, a uniform monolayer structure is ultimately created and the yield of viable cells is increased while maintaining proliferative activity. In the case of hybrid cultivation, the installation ensures their long-term cultivation (more than 60 days) while retaining the ability to produce monoclonal

antibodies.

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Compiled by Y. Utkin

Editor V. Petrash Tehred O. Sopko Corrector G. Reshetnik

Order 3668/26 Circulation 490Subscription

VNISHI USSR State Committee

for inventions and discoveries 113035, Moscow, Zh-35, Rautpska nab., d.L / 5

Production and printing company, Uzhgorod, Projecto st., 4

Claims (3)

1. INSTALLATION FOR CULTIVATION OF CELLS OR HYBRIDES, including throwing a cultivator, a gas supply system and a collection of culture medium, characterized in that, in order to more evenly distribute cells or hybrids along the growth surface and improve mass transfer in the pericellular region, the cultivator is equipped with a hollow glass , inside of which are placed along the height of the partition, having openings alternately located in the central and peripheral parts for the fluid flow, and an external chamber for the contact of the fluid and gas, the upper part five of which is connected with the upper cup portion and the bottom - the cultivator, the upper chamber is extended _β ive $ arranged therein at least one plate having holes with a bead ^ mi gas passage. 81eg ^ r ^w ns ^ 2. Installation according to π.1, which requires that, in order to increase the duration of hybrid cultivation, the installation is equipped with a pneumatic valve connecting the cultivator with a gas supply system with an external chamber for contacting the liquid and gas, the latter being connected in the middle the peristaltic pump to a vessel with a nutrient medium. 3. Installation according to π. 1, characterized in that, when growing cells, the upper part of the glass is connected to the upper part of the external chamber for contacting liquid and gas by means of a peristaltic pump.