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Serum-free culture medium for VERO serum-free cell culture and corresponding virus production

A serum-free medium, cell culture technology, applied in cell culture active agents, microorganism-based methods, animal cells, etc., can solve the problems of not widely used, high production cost, easy apoptosis of cells, etc., to reduce production costs. , The product quality is stable, and the effect of increasing the proportion of apoptosis

Inactive Publication Date: 2019-12-31
山东巨山能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, most of the current commercial serum-free media have low versatility, are not widely used, are not universal, and have high production costs, which is not conducive to commercial development and application; at the same time, the number of VERO cells grown in serum-free media is small , and the late persistence is poor, and the cells are prone to apoptosis during continuous passage. Based on this, we propose a serum-free medium for VERO serum-free cell culture and corresponding virus production

Method used

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  • Serum-free culture medium for VERO serum-free cell culture and corresponding virus production
  • Serum-free culture medium for VERO serum-free cell culture and corresponding virus production
  • Serum-free culture medium for VERO serum-free cell culture and corresponding virus production

Examples

Experimental program
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Effect test

Embodiment 1

[0022]A serum-free medium for VERO serum-free cell culture and corresponding virus production, including the following components: glycine 20mg / L, alanine 5mg / L, arginine hydrochloride 300mg / L, asparagine monohydrate 60mg / L, aspartic acid 10mg / L, cysteine ​​hydrochloride monohydrate 20mg / L, cystine dihydrochloride 75mg / L, glutamic acid 10mg / L, glutamine 200mg / L, histamine hydrochloride monohydrate Acid 40mg / L, Isoleucine 75mg / L, Leucine 75mg / L, Lysine Hydrochloride 95mg / L, Methionine 75mg / L, Phenylalanine 40mg / L, Proline 20mg / L , Serine 180mg / L, Threonine 60mg / L, Tryptophan 10mg / L, Tyrosine Disodium Salt Dihydrate 300mg / L, Valine 60mg / L, Hydroxyproline 15mg / L, Ornithine acid 60mg / L, taurine 75mg / L, human transferrin 15mg / L, recombinant insulin 10mg / L, 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) 4000mg / L, KERRY ultrafiltration plant peptone (kerry4601N ) 500mg / L, Angel yeast powder (Angel ultra) 500mg / L, ascorbic acid 60mg / L, sodium ascorbyl phosphonate 5mg / L, biotin ...

Embodiment 2

[0025] A serum-free medium for VERO serum-free cell culture and corresponding virus production, including the following components: glycine 15mg / L, alanine 2mg / L, arginine hydrochloride 150mg / L, asparagine monohydrate 20mg / L, aspartic acid 5mg / L, cysteine ​​hydrochloride monohydrate 15mg / L, cystine dihydrochloride 25mg / L, glutamic acid 5mg / L, glutamine 100mg / L, histamine hydrochloride monohydrate Acid 20mg / L, Isoleucine 25mg / L, Leucine 25mg / L, Lysine Hydrochloride 60mg / L, Methionine 50mg / L, Phenylalanine 20mg / L, Proline 15mg / L , Serine 150mg / L, Threonine 40mg / L, Tryptophan 5mg / L, Tyrosine Disodium Salt Dihydrate 200mg / L, Valine 25mg / L, Hydroxyproline 10mg / L, Ornithine acid 40mg / L, taurine 50mg / L, human transferrin 10mg / L, recombinant insulin 5mg / L, 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) 2000mg / L, KERRY ultrafiltration plant peptone (kerry4601N ) 100mg / L, Angel yeast powder (Angel ultra) 100mg / L, ascorbic acid 40mg / L, sodium ascorbyl phosphonate 1mg / L, biotin 0.1...

Embodiment 3

[0028] A serum-free medium for VERO serum-free cell culture and corresponding virus production, including the following components: glycine 18.75mg / L, alanine 4.45mg / L, arginine hydrochloride 247.5mg / L, monohydrate Tianmen Paragine 57.5mg / L, Aspartic Acid 6.65mg / L, Cysteine ​​Hydrochloride Monohydrate 17.56mg / L, Cystine Dihydrochloride 46.29mg / L, Glutamic Acid 7.35mg / L, Glutamine 190mg / L, histidine hydrochloride monohydrate 31.48mg / L, isoleucine 54.47mg / L, leucine 59.05mg / L, lysine hydrochloride 91.25mg / L, methionine 67.24mg / L, benzene Alanine 35.48mg / L, Proline 17.25mg / L, Serine 176.25mg / L, Threonine 53.45mg / L, Tryptophan 9.02mg / L, Tyrosine disodium salt dihydrate 275.79mg / L L. Valine 52.85mg / L, Hydroxyproline 13.1mg / L, Ornithine 50.58mg / L, Taurine 62.5mg / L, Ascorbic Acid 50mg / L, Sodium Ascorbyl Phosphonate 2.5mg / L, Biotin 0.5mg / L, choline chloride 8.98mg / L, D-calcium pantothenate 6.24mg / L, folic acid 2.65mg / L, niacin 2.02mg / L, pyridoxal hydrochloride 2.013mg / L, riboflavin ...

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Abstract

The invention discloses a serum-free culture medium for VERO serum-free cell culture and corresponding virus production. The culture medium comprises the following components: amino acids, vitamins, inorganic salts, auxiliary components and proteins. The amino acids comprise glycine, alanine, arginine hydrochloride, cystine dihydrochloride, glutamic acid, glutamine, histidine hydrochloride monohydrate, isoleucine, leucine, lysine hydrochloride, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine disodium salt dihydrate, valine, hydroxyproline, ornithine and taurine. Theproteins comprise human transferrin and recombinant insulin. When being used for VERO cell culture, the culture medium does not depend on the use of animal serum, does not introduce animal-derived protein, greatly lowers the production cost, sufficiently ensures the stable product quality, and can implement long-time culture and passage without increasing the apoptosis ratio.

Description

technical field [0001] The invention relates to the field of culture medium, in particular to a serum-free culture medium for VERO serum-free cell culture and corresponding virus production. Background technique [0002] Serum-free culture of VERO cells to produce vaccines has become the current mainstream trend. In line with the adherence characteristics of VERO cells and increased cell density, the design and optimization of serum-free chemically defined medium is the key to the serum-free culture of VERO cells. [0003] However, most of the current commercial serum-free media have low versatility, are not widely used, are not universal, and have high production costs, which is not conducive to commercial development and application; at the same time, the number of VERO cells grown in serum-free media is small , and the late persistence is poor, and the cells are prone to apoptosis during continuous passage. Based on this, we propose a serum-free medium for VERO serum-free...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00C12R1/93
CPCC12N5/0686C12N7/00C12N2500/32C12N2500/38C12N2501/998C12N2500/12C12N2500/34C12N2500/40C12N2500/46C12N2501/33C12N2500/16C12N2501/11C12N2500/14C12N2500/24
Inventor 齐智孟丹丹房圆瑗刘海英
Owner 山东巨山能源科技有限公司
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