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Serum-free medium for VERO cells

A serum-free culture medium and cell technology, applied in the field of culture medium, can solve the problems of poor cell adherence, unknown composition, and unsatisfactory cell proliferation.

Pending Publication Date: 2020-10-30
百奥特生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are some commercially available serum-free media, but these serum-free media have defects such as poor cell adhesion, unsatisfactory cell proliferation, high price, and unknown components.

Method used

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  • Serum-free medium for VERO cells
  • Serum-free medium for VERO cells
  • Serum-free medium for VERO cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A serum-free medium for VERO cells, including the following components: glycine 20mg / L, alanine 5mg / L, arginine 300mg / L, asparagine 59mg / L, aspartic acid 10mg / L , cysteine ​​20mg / L, cystine 75mg / L, glutamic acid 10mg / L, glutamine 200mg / L, histidine 40mg / L, phenylisoleucine 75mg / L, leucine 75mg / L, Lysine 94mg / L, Methionine 75mg / L, Phenylalanine 40mg / L, Proline 20mg / L, Serine 180mg / L, Threonine 59mg / L, Tryptophan 10mg / L, Tyrosine amino acid 300mg / L, valine 59mg / L, transferrin 14mg / L, pepsin 10mg / L, catalase 4000mg / L, ultra-filtered plant-derived peptone 480mg / L, yeast extract Extract 480mg / L, Vitamin C59mg / L, Vitamin C Sodium Phosphate 5mg / L, Vitamin H0.5mg / L, Choline 10mg / L, D-Calcium 10mg / L, Pteroylglutamate 5mg / L , anti-pyridosis factor 5mg / L, pyridoxal hydrochloride 5mg / L, riboflavin 0.5mg / L, cobalamin 5mg / L, inositol 14mg / L, vitamin K3 0.2mg / L, anhydrous Calcium nitrate 200mg / L, anhydrous magnesium sulfate 98mg / L, potassium chloride 400mg / L, dipotassium hydrogensu...

Embodiment 2

[0021]A serum-free medium for VERO cells, including the following components: glycine 14mg / L, alanine 2mg / L, arginine 140mg / L, asparagine 20mg / L, aspartic acid 5mg / L , cysteine ​​14mg / L, cystine 24mg / L, glutamic acid 5mg / L, glutamine 98mg / L, histidine 20mg / L, phenylisoleucine 24mg / L, leucine 24mg / L, Lysine 59mg / L, Methionine 50mg / L, Phenylalanine 20mg / L, Proline 14mg / L, Serine 140mg / L, Threonine 40mg / L, Tryptophan 5mg / L, Tyrosine amino acid 200mg / L, valine 24mg / L, transferrin 10mg / L, pepsin 5mg / L, catalase 2000mg / L, ultra-filtered plant-derived peptone 98mg / L, yeast extract Extract 98mg / L, Vitamin C 40mg / L, Vitamin C Sodium Phosphate 1mg / L, Vitamin H 0.1mg / L, Choline 5mg / L, D-Calcium 5mg / L, Pteroylglutamate 1mg / L , anti-pyridosis factor 1mg / L, pyridoxal hydrochloride 1mg / L, riboflavin 0.1mg / L, cobalamin 1mg / L, inositol 10mg / L, vitamin K3 0.05mg / L, anhydrous Calcium nitrate 140mg / L, anhydrous magnesium sulfate 50mg / L, potassium chloride 200mg / L, dipotassium hydrogen sulfate ...

Embodiment 3

[0023] A serum-free medium for VERO cells, including the following components: Glycine 17.37mg / L, Alanine 3.67mg / L, Arginine 248.1mg / L, Asparagine 57.4mg / L, Asparagine Acid 7.54mg / L, cysteine ​​18.14mg / L, cystine 45.67mg / L, glutamic acid 6.59mg / L, glutamine 154mg / L, histidine 30.54mg / L, phenylisoleucine Acid 49.46mg / L, Leucine 58.34mg / L, Lysine 91.24mg / L, Methionine 58.34mg / L, Phenylalanine 35.37mg / L, Proline 17.24mg / L, Serine 176.24mg / L , Threonine 52.67mg / L, Tryptophan 9.02mg / L, Tyrosine 275.79mg / L, Valine 52.85mg / L, Vitamin C 50mg / L, Vitamin C Sodium Phosphate 2.5mg / L, Vitamin H0 .5mg / L, Choline 8.24mg / L, D-Calcium 6.24mg / L, Pteroylglutamic Acid 1.67mg / L, Anti-Panacea Factor 2.02mg / L, Pyridoxal Hydrochloride 2.012mg / L, riboflavin 0.219mg / L, cobalamin 2.17mg / L, inositol 12.6mg / L, vitamin K3 0.1mg / L, anhydrous calcium nitrate 170mg / L, anhydrous magnesium sulfate 97.67mg / L, Potassium chloride 311.8mg / L, dipotassium hydrogensulfate 1200mg / L, sodium chloride 5800mg / L, sodium ...

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Abstract

The invention relates to a serum-free culture medium for VERO cells, which is characterized by comprising amino acids, proteins, vitamins, inorganic salts, serum replacement factors and auxiliary components, wherein the amino acids comprise glycine, alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, histidine, phenylisoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine L, the protein comprises transferrin, pepsase, catalase, ultrafiltration plant source peptone and yeast extract, and the serumreplacement factor comprises insulin, interleukin-2, an adhesion factor, a fibroblast growth factor, cortisol and thyroid hormone. According to the serum-free medium, rapid adherence of Vero cells can be promoted, the cultured Vero cells can keep a good cell form, the cell proliferation rate is higher, the components of the serum-free medium are simple, and the cost is reduced.

Description

technical field [0001] The invention relates to the field of culture medium, in particular to a serum-free culture medium for VERO cells. Background technique [0002] The role of cell culture in vaccine production is gaining more and more attention. In the past, in the process of culturing cells with serum, the uncertainty of the safety of cell expression products was increased, and the use of serum-containing medium for cell culture may lead to allergic reactions and batch-to-batch differences due to the presence of serum. Large size, easy contamination and high cost, this practical problem has become the driving force for the development of animal cell serum-free culture technology research and application. With the continuous advancement of serum-free culture technology, serum-free culture has become a general trend in the production of biotech drugs including vaccines. [0003] Vero cells are sensitive to a variety of viruses and are the most important cell substrate ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2500/90C12N2500/32C12N2500/25C12N2501/734C12N2501/71C12N2501/998C12N2500/74C12N2501/2302C12N2501/58C12N2501/115C12N2501/395C12N2501/39C12N2500/14C12N2500/16C12N2500/12C12N2500/20C12N2500/24C12N2500/22C12N2500/34C12N2500/46C12N2500/30C12N2500/40C12N2501/11C12N2501/105C12N2500/36C12N2501/80
Inventor 唐贺文孙正宇
Owner 百奥特生物科技(上海)有限公司
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