Low-serum medium suitable for growth of Vero cells

A low serum and culture medium technology, applied in the field of cell culture without serum or low serum, can solve problems such as poor safety, and achieve the effect of reducing pollution, clear source, and promoting growth

Inactive Publication Date: 2013-06-19
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to overcome the disadvantages that high content of calf serum must be used to promote growth and poor safety in the conventional culture of Vero cells, and provides a new low-serum medium

Method used

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

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Embodiment 1

[0010] The invention is composed of 5% calf serum, soybean plant hydrolyzed protein (component V) 5g / L, epidermal growth factor 20μg / L, glutamine 2mM and beta-mercaptoethanol 30μM.

[0011] 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

[0013] The present 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.

[0014] 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.

[0015] Preparation method:

[0016] 1) Preparation of soybean plant hydrolyzed protein stock solution When using soybean plant hydrolyzed protein for basic growth culture on a laboratory scale, it is necessary to use a concentrated and sterilized stock solution. In our laboratory, the concentration of 100g / L is used as the storage solution. Weigh 10 g of soybean plant hydrolyzed protein powder, dissolve it i...

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Abstract

The invention develops a low-serum medium suitable for growth of Vero cells. An amount of calf serum in conventional culturing is reduced from 10 % (V component) to 2 %, and conventionally added animal source components are replaced by plant source components and recombinant protein. The low-serum medium can reduce amount of serum, and thus not only reduces cost, but also minimizes possibilities that the serum in the medium carries exogenous pathogen contamination and reduces a lot of other proteins in the serum, thereby benefiting separation and purification of downstream products. By using a plant protein hydrolyzate and a recombinant epidermal growth factor to replace animal source components, side effects of animal source components can be reduced, and safety of biological products can be improved. The medium provided by the invention not only enables the Vero cells to grow well, but also reduces cost, minimizes exogenous pathogen contamination, and improves production stability of cell products, thereby providing a high-quality medium for producing of biological products with the Vero cells as virus host or expression carrier.

Description

1. 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. 2. Background technology [0002] Vero cell is a cell line approved by the World Health Organization and my country's biological products regulations for the production of vaccines. It has the advantages of convenient source, high biological safety, sensitivity to infection by various viruses, and high virus proliferation titer. Calf serum with a final concentration of 10% (volume) was used in common 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 numbe...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 汤承岳华苏晓蕊
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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