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Serum-free medium used for culturing Vero cells, and preparation method thereof

A technology of serum-free medium and culture medium, which is applied in the field of serum-free medium and its preparation, can solve the problems of poor cell adhesion, complex components, and unsatisfactory cell proliferation rate, etc., and achieve simple components and simple preparation methods , the effect of excellent cell performance

Inactive Publication Date: 2016-03-30
ZHAOQING DAHUANONG BIOLOGIC PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the prior art has problems such as poor cell adhesion, too complex components, and unsatisfactory cell proliferation rate.

Method used

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  • Serum-free medium used for culturing Vero cells, and preparation method thereof
  • Serum-free medium used for culturing Vero cells, and preparation method thereof
  • Serum-free medium used for culturing Vero cells, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A serum-free medium for cultivating Vero cells consists of the following components by volume percentage: 90% of basal culture solution, 4% of amino acid solution, 5% of serum replacement factor solution, 0.1% of antioxidant solution, and yeast extract solution 0.8%, ethanolamine solution 0.1%;

[0041] The solvent in the amino acid solution is the basic culture solution, and the solute is composed of the following components by weight: 0.5 g of lysine, 1 g of tryptophan, 0.5 g of phenylisoleucine, 1 g of valine, 1 g of leucine, and 1 g of glycine 0.5g, Isoleucine 1g, Threonine 0.5g, Serine 0.5g, Arginine 1g, Aspartic Acid 0.5g and Glutamine 0.5g;

[0042] The solute of the serum replacement factor is a mixture obtained by mixing cortisol, transferrin and protamine in a ratio of 0.5:0.5:0.8;

[0043] The solvent of the antioxidant solution is basal culture fluid, the solute is glutathione, and the concentration is 1mg / L;

[0044] The concentration of the yeast extract...

Embodiment 2

[0053] A serum-free medium for cultivating Vero cells consists of the following components by volume percentage: 92% of basal culture solution, 2% of amino acid solution, 5% of serum replacement factor solution, 0.2% of antioxidant solution, and yeast extract solution 0.7%, ethanolamine solution 0.1%;

[0054] The solvent in the amino acid solution is the basic culture solution, and the solute is composed of the following components by weight: lysine 1g, tryptophan 0.5g, phenylisoleucine 1g, valine 0.5g, leucine 0.5g, Glycine 1g, Isoleucine 1g, Threonine 0.5g, Serine 1g, Arginine 0.5g, Aspartic Acid 0.5g and Glutamine 0.5g;

[0055] The solute of the serum replacement factor is a mixture obtained by mixing insulin, chromatin and polylysine in a ratio of 0.4:0.2:0.6;

[0056] The solvent of the antioxidant solution is a basal culture solution, the solute is a carotenoid, and the concentration is 0.5mg / L;

[0057] The concentration of the yeast extract solution is 8mg / L;

[0...

Embodiment 3

[0061] A serum-free medium for cultivating Vero cells consists of the following components by volume percentage: 92% of basal culture solution, 3% of amino acid solution, 4% of serum replacement factor solution, 0.2% of antioxidant solution, and yeast extract solution 0.6%, ethanolamine solution 0.2%;

[0062] The solvent in the amino acid solution is the basal culture solution, and the solute is composed of the following components by weight: 0.5 g of lysine, 1 g of tryptophan, 1 g of phenylisoleucine, 1 g of valine, 1 g of leucine, 0.5 g of glycine g, isoleucine 1g, threonine 0.5g, serine 1g, arginine 1g, aspartic acid 0.5g and glutamine 1g;

[0063] The solute of the serum replacement factor is a mixture obtained by mixing epidermal growth factor, phosphoprotein and lipoprotein in a ratio of 0.6:0.4:0.8;

[0064] The solvent of the antioxidant solution is basal culture fluid, the solute is citric acid, and the concentration is 0.8mg / L;

[0065] The concentration of the ye...

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Abstract

The invention relates to a serum-free medium used for culturing Vero cells. The serum-free medium is composed of, by volume, 85 to 95% of a base culture solution, 2 to 5% of an amino acid solution, 2 to 7% of a serum alternative factor solution, 0.1 to 0.5% of an antioxidant solution, 0.8 to 2% of a yeast extract solution, and 0.1 to 0.5% of an ethanol amine solution. The invention also provides a preparation method of the serum-free medium. In the preparation method, the base culture solution, the amino acid solution, the serum alternative factor solution, the antioxidant solution, the yeast extract solution, and the ethanol amine solution are mixed at the above ratio so as to obtain the serum-free medium. The serum-free medium used for culturing Vero cells is capable of promoting rapid attachment of Vero cells; cell morphology of the Vero cells cultured with the serum-free medium can be maintainer preferably, and cell proliferation rate is higher; the composition of the serum-free medium is simple; cost is reduced; and preparation is convenient.

Description

technical field [0001] The invention belongs to the field of biological products, and relates to a serum-free medium for cultivating Vero cells and a preparation method thereof. Background technique [0002] Since WHO called for the use of cells as substrates to produce vaccines in 1995, people have paid more and more attention to the role of cell culture in vaccine production. 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 biote...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0602
Inventor 陈瑞爱徐家华唐兆新张文炎高艳
Owner ZHAOQING DAHUANONG BIOLOGIC PHARMA
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