Culture medium used for Vero cell and cultivation method thereof

A medium and serum-free medium technology, applied in the field of biological products, can solve the problems of complex purification process, large differences between product batches, high cost and so on

Inactive Publication Date: 2008-05-21
天津百若克医药生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Animal serum is a culture medium and a commonly used substance in the field of biological products. It is very necessary in the culture of bacteria and cells, but serum has obvious and unreliable shortcomings: the composition is not clear, the difference between product batches is large, and it is easy to spread bacteria, viruses and other diseases , the price is expensive, and the downstream purification process of the recombinant protein produced by the cells cultured with serum is complicated and costly
The Vero cells used in the industrial production of recombinant proteins are characterized by adherent growth, and the area of ​​the attached substrate that the culture system can provide is limited, making it difficult to greatly increase the number of proliferation and expansion

Method used

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  • Culture medium used for Vero cell and cultivation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0341] The serum-free culture method of Vero cells, wherein, the Vero cells grown on the wall are adapted to the SFM serum-free medium through the Vero cell suspension to adapt to the culture, and the Vero growth mode can be converted from the wall-attached growth to the suspension growth; further, the Vero cells can be grown in Vero expands growth in CHEMICALLY DEFINED MEDIUM (chemically defined synthetic medium) medium. The culture conditions are as follows: 37 °C, 5% CO 2 , a culture environment with a humidity greater than 95%.

Embodiment 2

[0343] The culture method is as described in Example 1, wherein the components and ratios of Vero cell suspension adapted to SFM serum-free medium and Vero expansion CHEMICALLY DEFINED MEDIUM medium are as follows:

[0344] Vero cell suspension adapted to SFM serum-free medium composition and ratio:

[0345] Composition mg / L

[0346] Glycine 33.5

[0347] L-Alanine (L-Alanine) 16.78

[0348] L-arginine (L-Arginine) 124.2

[0349] L-Asparagine (L-Asparagine) 31.002

[0350] L-Aspartic acid (L-Aspartic acid) 6.66

[0351] Cysteine ​​hydrochloride-water (L-Cysteine ​​hydrochloride-H 2 O) 0.06

[0352] L-Cystine dihydrochloride (L-Cystine 2HCl) 35.624

[0353] L-Glutamic Acid 51.94

[0354] L-glutamine (L-Glutamine) 149.2

[0355] L-Histidine 20.328

[0356] L-Hydroxyproline (L-Hydroxyproline) 10

[0357] L-Isoleucine 34.8

[0358] L-Leucine (L-Leucine) 48.62

[0359] L-Lysine hydrochloride (L-Lysine hydrochloride) 57.3

[0360] L-Methionine 12.9

[0361] L-Phenylal...

Embodiment 3

[0487] The culture method is as described in Example 1, wherein the components and ratios of Vero cell suspension adapted to SFM serum-free medium and Vero expansion CHEMICALLY DEFINED MEDIUM medium are as follows:

[0488] Vero cell suspension adapted to SFM serum-free medium composition and ratio:

[0489] Composition mg / L

[0490] Glycine 50

[0491] L-Alanine 5

[0492] L-Arginine (L-Arginine) 200

[0493] L-Asparagine 10

[0494] L-Aspartic acid (L-Aspartic acid) 15

[0495] L-cysteine ​​hydrochloride (L-Cysteine ​​hydrochloride-H 2 O) 0.01

[0496] L-Cystine 2HCl 100

[0497] L-Glutamic Acid 10

[0498] L-Glutamine 200

[0499] L-Histidine 5

[0500] L-Hydroxyproline (L-Hydroxyproline) 20

[0501] L-Isoleucine 10

[0502] L-Leucine 100

[0503] L-Lysine hydrochloride 15

[0504] L-Methionine 30

[0505] L-Phenylalanine 5

[0506] L-Proline (L-Proline) 100

[0507] L-Serine (L-Serine) 5

[0508] L-Threonine 50

[0509] L-Tryptophan 2

[0510] L-Tyrosin...

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Abstract

The invention relates to the preparation and application method of a medium, which is specifically related to a medium which is used for Vero cell and a culture method of the Vero cell. The invention belongs to the biological product area. The culture method of the invention is that the adhering Vero cells are suspend and cultured in SFM serum-free medium for the domestication and adaptation; the growing pattern of the Vero cell changes from adherence growth to suspension growth; the Vero cells are further amplified in Vero amplification CHEMICALLY DEFINED MEDIUM. The method of the invention has good effects; the growing pattern of the Vero cell is transformed from adherence growth to suspension growth, which changes the growth pattern of the cell and greatly increases the yield of the cell.

Description

technical field [0001] The invention relates to a method for preparing and using a culture medium, more specifically to a culture medium for Vero cells and a culture method thereof. It belongs to the field of biological products. Background technique [0002] Animal serum is a culture medium and a commonly used substance in the field of biological products. It is very necessary in the culture of bacteria and cells, but serum has obvious and unreliable shortcomings: the composition is not clear, the difference between product batches is large, and it is easy to spread bacteria, viruses and other diseases , the price is expensive, and the downstream purification process of the recombinant protein produced by the cells cultured with serum is complicated and expensive. The Vero cells used in the industrial production of recombinant proteins are characterized by adherent growth, and the area of ​​the attached substrate that the culture system can provide is limited, making it di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/071
Inventor 黄宗堂唐贺文
Owner 天津百若克医药生物技术有限责任公司
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