Serum-free culture medium for culturing Vero cells and preparation method of serum-free culture medium

A serum-free medium and basal medium technology, applied in biochemical equipment and methods, culture process, tissue culture and other directions, can solve the problems of poor cell adherence, unknown composition, and unsatisfactory cell proliferation rate, and achieve low cost. , controllable between batches, and the effect of avoiding biosafety risks

Pending Publication Date: 2022-05-27
杭州荣泽生物科技集团有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In vaccine production, in order to achieve high-density culture of cells, 5%-10% serum is usually added to the basal medium. Serum can provide various nutrients required for cell growth, such as hormones, trace elements, adherence Factors, etc., but there are many shortcomings in the use of serum. Therefore, the development of serum-free medium is an important condition for the clinical application of cells. First, some of the existing commercially available serum-free medium have poor cell adhesion and unsatisfactory cell proliferation rate. , unknown components and other defects, therefore, the present invention provides a serum-free medium for cultivating Vero cells and a preparation method thereof

Method used

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  • Serum-free culture medium for culturing Vero cells and preparation method of serum-free culture medium
  • Serum-free culture medium for culturing Vero cells and preparation method of serum-free culture medium
  • Serum-free culture medium for culturing Vero cells and preparation method of serum-free culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Medium 1: basal medium, 800 mg / L vegetable hydrolyzed protein, 2.8 mg / L Fe 3+ -Sulfate-iminodiacetic acid complex, 7.2mg / L Fe 3+ -Sulfate-glycylglycine complex, 5mg / L insulin, 1.28mg / L auxin, 1.0mg / L hydrocortisone, 2.2mg / L cortisol, 0.01mg / L epidermal growth factor EGF, 0.6mg / L transforming growth factor TGF, 0.15mg / L platelet growth factor PDGF, 1.8mg / L fibroblast growth factor FGF, 2.5mg / L fibronectin, 1.4mg / L collagen, 0.3mg / L laminin, 1.1mg / L poly-L-lysine, 0.4mg / L ornithine, 0.01mg / L sodium selenite, 0.03mg / L gold tricarboxylic acid, 0.008mg / L Zn 2+ , 0.07mg / L Riboflavin, 0.22mg / L Folic Acid, 0.17mg / L Cobalamin, 0.1mg / L Calcium Pantothenate, 7.68mg / L Inositol, 0.3mg / L Niacinamide, 3.5mg / L Chloride Choline, 0.04mg / L lipoic acid, 0.02mg / L pyridoxal hydrochloride, 0.27mg / L vitamin C, 0.01mg / L vitamin E, 0.23mg / L pyridoxine hydrochloride, 0.005mg / L carboxylesterase , 301.44mg / L potassium chloride, 13mg / L calcium chloride, 118mg / L sodium dihydrogen phosp...

Embodiment 2

[0054] Example 2: Medium 2: basal medium, 500mg / L serum albumin, 15mg / L transferrin, 5mg / L insulin, 1.28mg / L auxin, 1.0mg / L hydrocortisone, 2.2mg / L L cortisol, 0.01mg / L epidermal growth factor EGF, 0.6mg / L transforming growth factor TGF, 0.15mg / L platelet growth factor PDGF, 1.8mg / L fibroblast growth factor FGF, 2.5mg / L fibronectin, 1.4mg / L collagen, 0.3mg / L laminin, 1.1mg / L poly-L-lysine, 0.4mg / L ornithine, 0.01mg / L sodium selenite, 0.03mg / L gold essence Tricarboxylic acid, 0.008mg / L Zn 2+ , 0.07mg / L Riboflavin, 0.22mg / L Folic Acid, 0.17mg / L Cobalamin, 0.1mg / L Calcium Pantothenate, 7.68mg / L Inositol, 0.3mg / L Niacinamide, 3.5mg / L Chloride Choline, 0.04mg / L lipoic acid, 0.02mg / L pyridoxal hydrochloride, 0.27mg / L vitamin C, 0.01mg / L vitamin E, 0.23mg / L pyridoxine hydrochloride, 0.005mg / L carboxylesterase , 301.44mg / L potassium chloride, 13mg / L calcium chloride, 118mg / L sodium dihydrogen phosphate, 6801mg / L sodium chloride, 68.835mg / L magnesium sulfate, 0.15mg / L linoleic acid, ...

Embodiment 3

[0055] Example 3: Medium 3: basal medium, 500mg / L serum albumin, 15mg / L transferrin, 5mg / L insulin, 1.28mg / L auxin, 1.0mg / L hydrocortisone, 2.2mg / L L cortisol, 0.01mg / L epidermal growth factor EGF, 0.6mg / L transforming growth factor TGF, 0.15mg / L platelet growth factor PDGF, 1.8mg / L fibroblast growth factor FGF, 2.5mg / L fibronectin, 1.4mg / L collagen, 0.3mg / L laminin, 1.1mg / L poly-L-lysine, 0.4mg / L ornithine, 0.01mg / L sodium selenite, 0.03mg / L gold essence Tricarboxylic acid, 0.008mg / L Zn 2+ , 0.07mg / L Riboflavin, 0.22mg / L Folic Acid, 0.17mg / L Cobalamin, 0.1mg / L Calcium Pantothenate, 7.68mg / L Inositol, 0.3mg / L Niacinamide, 3.5mg / L Chloride Choline, 0.04mg / L lipoic acid, 0.02mg / L pyridoxal hydrochloride, 0.27mg / L vitamin C, 0.01mg / L vitamin E, 0.23mg / L pyridoxine hydrochloride, 0.005mg / L carboxylesterase , 301.44mg / L potassium chloride, 13mg / L calcium chloride, 118mg / L sodium dihydrogen phosphate, 6801mg / L sodium chloride, 68.835mg / L magnesium sulfate, 1.2mg / L amphotericin B, ...

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Abstract

The invention discloses a serum-free culture medium for culturing Vero cells and a preparation method thereof, the serum-free culture medium comprises a basic culture medium, the basic culture medium comprises glucose and glutamine, and the serum-free culture medium also comprises a main addition factor and a secondary addition factor, the main addition factors comprise proteins, hormones, growth factors, adherence promoting factors, trace elements and vitamins, and the secondary addition factors comprise a reducing agent, a trypsin inhibitor and a shear force protective agent. According to the culture medium, the Vero cells are cultured through the culture medium, normal growth of the Vero cells can be maintained, the amplification efficiency of the Vero cells can be improved, research, development and production of new coronavirus vaccines can be accelerated, the culture medium is clear in chemical component, free of serum and low in cost, biological safety risks caused by using serum are avoided, and the culture medium is suitable for popularization and application. The stability of a biological product production process and finished product quality is favorably improved.

Description

technical field [0001] The invention relates to the technical field of Vero cell culture medium, in particular to a serum-free medium for culturing Vero cells and a preparation method thereof. Background technique [0002] Vero cell is a vaccine production cell line approved by the World Health Organization and my country's biological product regulations. It has been used for the production of various vaccines such as rabies virus and influenza virus. In addition, Vero cells are also excellent host cells. [0003] In vaccine production, in order to achieve high-density culture of cells, 5%-10% serum is usually added to the basal medium. Serum can provide various nutrients required for cell growth, such as hormones, trace elements, adherence factors, etc., but the use of serum has many shortcomings. Therefore, the development of serum-free medium is an important condition for cells to move towards clinical application. First, some of the current commercially available serum-f...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2500/90C12N2501/998C12N2500/25C12N2501/39C12N2501/11C12N2501/135C12N2501/115C12N2533/52C12N2533/54C12N2533/32C12N2500/12C12N2500/22C12N2500/38C12N2500/35C12N2500/46C12N2500/30C12N2500/44C12N2500/14C12N2500/16C12N2500/36C12N2501/999C12N2500/60C12N2501/305C12N2501/335C12N2501/395C12N2501/392C12N2501/73C12N2501/70C12N2501/71C12N2501/148C12N2501/15
Inventor 陈相波雷鸣田朋飞应荣
Owner 杭州荣泽生物科技集团有限公司
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