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Method for scale-up culture of animal cells

A technology for animal cells and cell culture, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of low cell expansion culture efficiency, inconsistent cell growth status, large space occupation, etc., and achieve cell expansion culture efficiency High, reduce the unevenness of cell growth and save production space

Inactive Publication Date: 2014-06-04
BEIJING SKYWING TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the disadvantages of the existing method of using cell culture flasks for animal cell expansion and cultivation, such as large space occupation, cumbersome operation, low efficiency of cell expansion and cultivation, inconsistent cell growth conditions, etc., and to provide a new method for animal cell expansion and cultivation. The method uses a cell factory instead of a traditional cell culture bottle to carry out cell expansion before microcarrier culture in a bioreactor. The method is simple to operate, can increase the production scale per unit space, improve cell viability and cell recovery rate, thereby improving the expansion of cell culture. efficiency, and reduce the risk of contamination

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0069] Cell factory-120L bioreactor microcarrier culture Marc145 cells to produce PRRS vaccine

[0070] First, Marc145 cells cryopreserved in liquid nitrogen were resuscitated, then amplified and cultured sequentially through square bottles and spinner bottles, and finally expanded to 4 cells with a culture area of ​​850 cm 2 The free cell suspension was obtained by digestion in a spinner bottle of the same method, wherein the cell culture medium used was a modified DMEM cell culture medium (CatNo.MD210, Beijing Tsingda Tianyi Technology Co., Ltd.) suitable for Marc145 cell culture. The cell suspension obtained above was divided into two equal parts, one part was used for the following experiments, and the other part was used for Comparative Example 1.

[0071] Then, follow the steps below to perform scale-up culture of Marc145 cells in the cell factory.

[0072] 1) Culture Marc145 cells in a 10-layer cell factory

[0073] A 10-layer cell factory (1 CF10, manufactured by the...

Embodiment 2-4

[0082] According to the conditions recorded in Table 1 and the same operations as in Example 1, Examples 2-4 were carried out to obtain large-scale cultured animal cells that can be used in the production of downstream biological products.

[0083] Table 1

[0084]

[0085]

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Abstract

The invention relates to a method for scale-up culture of animal cells. The method includes the following steps of firstly, culturing animal cells in a cell factory; secondly, digesting the cultured animal cells, namely removing culture solution, and washing the cultured animal cells by pH7.4-8.0 washing solution; removing the washing solution by adding pH7.4-8.0 digestive solution 20-80mL per layer, allowing the digestive solution to evenly flow to each layer to digest the cells at 36-38 DEG C, removing partial digestive solution when more than 60% of cells fall off from the cell culture surface, and continuing to digest by the residual 10-35% of digestive solution; and adding fresh cell culture solution containing serum or digestive solution inhibitor to finish digesting and performing cell suspension again, and adding the culture solution for once to three times repeatedly; and thirdly, inoculating obtained cell suspension to a bioreactor for culture.

Description

technical field [0001] The invention relates to a method for amplified cultivation of animal cells, in particular to a method for amplified cultivation of animal cells with a cell factory. Background technique [0002] Large-scale animal cell culture technology refers to the technology of cultivating animal cells at high density and in large quantities in cell culture bioreactors under artificial conditions (setting pH, temperature, dissolved oxygen, etc.) for the production of biological products. It is widely used in the research and development and industrial production of biological products such as antibodies and virus vaccines, and is the core technology in the production of biological products. [0003] In the actual production process, before large-scale cultivation of animal cells to produce target biological products, the number of cells that meet the production requirements must first be obtained by expanding the seed cells. The commonly used amplification method...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/07C12N5/071
Inventor 王建超高飞陈文庆张韧
Owner BEIJING SKYWING TECH CO LTD
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