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Liquid change method for bioreactor and bioreactor using method

A technology of bioreactors and organisms, applied in biochemical equipment and methods, biochemical instruments, tissue cell/virus culture devices, etc., can solve the problem of large damage to cultured products, large space occupied by motors, complicated installation and disassembly operations, etc. problem, achieve the effect of reducing mechanical damage and cost

Inactive Publication Date: 2010-10-27
BEIJING SKYWING TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] The technical problem to be solved by the present invention is to overcome the large damage to the cultured species and high production cost of the current liquid exchange method used in bioreactors, and the complicated installation and disassembly operation of the bioreactor using the liquid exchange method of the prior art, which cannot be cleaned online, Driven by the agitator shaft of the culture tank, it has the disadvantages of difficult sealing of the culture tank, large space occupied by the motor and high production cost; it provides a liquid replacement method for bioreactors with little damage to the culture and low production cost and uses the existing technology Bioreactor for liquid exchange method

Method used

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  • Liquid change method for bioreactor and bioreactor using method
  • Liquid change method for bioreactor and bioreactor using method
  • Liquid change method for bioreactor and bioreactor using method

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Bioreactor: Clavorus 500L bioreactor for animal cell culture (manufactured by Beijing Tianherui Biotechnology Co., Ltd.). This instrument is a kind of existing bioreactor, and the bioreactor has a rotary filter. When using the method of the present invention to change the liquid, stop the rotation of the stirring shaft.

[0060] Culture: BHK21 cells

[0061] Medium: BHK21 cell suspension medium

[0062] (MD910, manufactured by Beijing Qingda Tianyi Biotechnology Co., Ltd.)

[0063] Cell culture process:

[0064] The volume of cultured cells and medium contained in the bioreactor is 420L, when BHK21 cells grow to 3×10 6 cells / ml, stop the rotation of the stirring shaft, and after standing for 30 hours, 90% of the cells have settled down to below one-third of the height of the medium liquid level, and the height of the medium liquid level refers to the height of the medium liquid level to The height of the bottom of the bioreactor, that is, the cells are concentrated ...

Embodiment 2

[0066] Bioreactor: Clavorus 2000L animal cell culture bioreactor (manufactured by Beijing Tianherui Biotechnology Co., Ltd.). When using the method of the present invention to change liquid, stop stirring.

[0067] Culture: CHO cells

[0068] Medium: Serum-free animal component-free medium for CHO cells

[0069] (SAF01, manufactured by Beijing Qingda Tianyi Biotechnology Co., Ltd.)

[0070] Cell culture process:

[0071] The volume of cultured cells and medium contained in the bioreactor is 1500L, when BHK21 cells grow to 3.3×10 6 cells / ml, stop the rotation of the stirring shaft, and after standing for 20 hours, 80% of the cells have settled down to below 1 / 2 of the height of the medium liquid level. The height of the bottom of the bioreactor. Raise the pressure in the bioreactor to 0.1MPa higher than the ambient pressure, remove the upper 700L medium; reduce the pressure in the bioreactor to the ambient pressure, and inject fresh medium to a volume of 1500L. After changi...

Embodiment 3

[0073] Bioreactor: Clavorus 500L bioreactor for animal cell culture (manufactured by Beijing Tianherui Biotechnology Co., Ltd.). This instrument is a kind of existing bioreactor, and the bioreactor has a rotary filter. The reactor was modified as follows: the spin filter was removed and the collection tube opening was located 8 cm from the bottom of the bioreactor, as figure 1 shown.

[0074] Culture: BHK21 cells

[0075] Medium: BHK21 cell suspension medium

[0076] (MD910, manufactured by Beijing Qingda Tianyi Biotechnology Co., Ltd.)

[0077] Cell culture process:

[0078] The volume of cultured cells and medium contained in the bioreactor is 420L, when BHK21 cells grow to 3×10 6 When cells / ml, stop the stirring shaft to rotate, after standing for 30 hours, take a sample and find that about 90% of the cells have settled down to below 5cm from the bottom of the reactor, use sterile compressed air to raise the pressure in the bioreactor to 0.03MPa, remove 380L medium in...

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Abstract

The invention provides a liquid change method for a bioreactor, which comprises the following steps of: 1) standing the first culture liquid containing the cultured substance until at least 50 percent of the cultured substance is settled below 1 / 2 part of the liquid level height of the first culture liquid, wherein the liquid level height of the culture liquid refers to the height from the liquid level of the first culture liquid to the bottom of the bioreactor; 2) moving out the first culture liquid with at least 1 / 2 volume on the upper part; and 3) injecting the second culture liquid. The invention also provides a bioreactor which is applied to the liquid change method. Because the liquid change method does not comprise the step of centrifugal filtration, the mechanical damage to the physiological structure of the cultured substance due to centrifugal filtration shear force is reduced, and the cost is reduced. The bioreactor applied to the method also has the characteristics of simple operation, high flexibility, easily control and setting of the liquid change ratio in advance, long continuous work period, high sealing performance, and hard pollution.

Description

technical field [0001] The invention relates to a liquid replacement method for liquid suspension culture of biological materials and culture equipment using the method. More specifically, it relates to a liquid replacement method for a bioreactor and a bioreactor using the method. Background technique [0002] In organisms (such as various microorganisms such as bacteria, fungi, viruses, etc.), isolated organs of organisms (such as root tips, shoot tips, cotyledons, leaf primordia, flower primordia, etc.) of organisms, isolated tissues of organisms ( For example, in the liquid culture process of plant callus, etc.) or organism cells (such as various animal cells), it is necessary to regularly replace the culture liquid (medium, etc.) to ensure that sufficient nutrients are supplied to the cultured, and timely Discharge of the metabolic waste of the culture. The process of exchanging the culture fluid is called fluid exchange. Therefore, the requirements for the bioreacto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/02
Inventor 张韧陈文庆王建超刘俊生
Owner BEIJING SKYWING TECH CO LTD