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Low-serum culture medium suitable for growth of Vero cells

A low serum and medium technology, applied in the field of cell culture without serum or low serum, can solve problems such as poor safety, and achieve the effects of reducing pollution, good safety and uniform composition

Inactive Publication Date: 2018-06-01
宁波高新区绿元汇生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to overcome the disadvantages that high content of calf serum must be used in the conventional culture of Vero cells to promote growth and poor safety, and provides a new low-serum medium

Method used

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  • Low-serum culture medium suitable for growth of Vero cells
  • Low-serum culture medium suitable for growth of Vero cells
  • Low-serum culture medium suitable for growth of Vero cells

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Embodiment 1

[0011] The invention is composed of 5% calf serum, soybean plant hydrolyzed protein (component V) 5g / L, epidermal growth factor 20μg / L, glutamine 2mM and β-mercaptoethanol 30μM.

[0012] This embodiment reduces the amount of calf serum used in conventional culture, uses soybean plant hydrolyzed protein, and its cost is lower than that of conventional medium, and at the same time solves the problem of premature cell death caused by reducing the use of calf serum. problem of undersaturation.

Embodiment 2

[0014] The invention consists of 2% calf serum, soybean plant hydrolyzed protein (V component) 5g / L, epidermal growth factor 60μg / L, glutamine 4mM, sodium selenite 50nM, ferric citrate 100μM, vitamin C 100μM, ethanolamine 200 μM composition, β-mercaptoethanol 30 μM, sodium pyruvate 1 mM.

[0015] In this embodiment, the amount of serum is further reduced, plant hydrolyzed protein and epidermal growth factor are increased, the density of cells is increased, and the specific growth rate is increased, thereby increasing the vaccine production in the medical field.

[0016] Preparation method:

[0017] Preparation of Soy Plant Protein Hydrolyzate Stock Solutions Using soy plant protein hydrolyzates for basic growth cultures at laboratory scale requires the use of concentrated and sterile stock solutions. In our laboratory, the concentration of 100g / L is used as the stock solution. Weigh 10g of soybean plant hydrolyzed protein powder, dissolve it in 100ml of basal medium, and ste...

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Abstract

The invention provides a low-serum culture medium suitable for the growth of Vero cells. The dosage of calf serum for conventional culture is reduced to 2 percent from 10 percent (which is the contentof a component V), and conventionally added animal derived components are replaced with plant derived components and recombinant protein. According to the low-serum culture medium provided by the invention, the dosage of the serum is reduced, so that the cost is reduced, and the possibility that the serum in the culture medium carries exogenous pathogenic bacterium pollution is reduced; meanwhile, a lot of hybrid proteins in the serum are reduced and separation and purification of down-stream products are facilitated; plant hydrolyzed protein and recombinant endothelial growth factors are used for replacing the animal derived components so that the side effect of the animal derived components is reduced and the safety of biological products is improved. According to the culture medium provided by the invention, the Vero cells grow well and the cost is reduced; the exogenous pathogenic bacterium pollution is reduced, a post-period purification technology is reduced and the stability ofproducing cell products is improved; one good-quality culture medium is provided for producing the biological products which take the Vero cells as virus hosts or expression vectors.

Description

technical field [0001] The invention relates to the technical field of serum-free or low-serum culture of cells, in particular to a low-serum medium suitable for the growth of Vero cells, which is used for the low-serum culture of Vero cells and the proliferation of viruses. Background technique [0002] The cell is a cell line for vaccine production approved by the World Health Organization and my country's biological products regulations. It has the advantages of convenient source, high biological safety, sensitivity to infection by various viruses, and high virus proliferation titer. Calf serum was used at a final concentration of 10% (volume) in ordinary Vero cell culture medium. However, the chemical composition contained in the serum is uncertain, and there are differences between each batch. At the same time, the price of the serum is expensive, and it is easy to cause contamination by bacteria, fungi, viruses and mycoplasma; in addition, a large number of complex pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2500/12C12N2500/24C12N2500/32C12N2500/38C12N2500/44C12N2500/46C12N2501/11C12N2501/998
Inventor 袁十义
Owner 宁波高新区绿元汇生物科技有限公司
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