Low-serum protein-free culture medium applicable to Vero cell growth and preparation method thereof

A protein-free medium and low-serum technology, applied in the field of low-serum protein-free medium and its preparation, can solve the problems of standardization and serialization limitations, unclear chemical composition, expensive serum, etc., to achieve easy pollution and benefit The effect of separation and chemical composition

Inactive Publication Date: 2015-12-23
南京三生生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the chemical composition contained in the serum is not clear, and there are quality differences between each batch, which limits the standardization and continuity of the experiment, and is likely to cause biological contamination such as bacteria, fungi, viruses, and mycoplasma. A large number of complex proteins will bring great difficulties to the separation and purification of biological products such as vaccines, cytokines, and monoclonal antibodies. More importantly, serum is expensive and difficult to source

Method used

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  • Low-serum protein-free culture medium applicable to Vero cell growth and preparation method thereof
  • Low-serum protein-free culture medium applicable to Vero cell growth and preparation method thereof
  • Low-serum protein-free culture medium applicable to Vero cell growth and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] This example is used to prepare a low-serum, protein-free cell culture medium suitable for Vero cell growth.

[0042] 1. The formulation of the culture medium

[0043] Table 1 medium formula

[0044]

[0045]

[0046]

[0047] 2. Preparation method

[0048] (1) Get the dried components in the formula and put them into the ball mill tank respectively, and ball mill for 2 hours to get the culture medium dry powder;

[0049] (2) Dissolve 16.73 g of the medium dry powder prepared in step (1) in 900 ml ultrapure water, then dilute it to 1 L with ultrapure water, and filter and sterilize it with a 0.22 μm filter membrane to obtain suitable Low-serum, protein-free liquid medium grown on Vero cells, refrigerated at 6°C for later use.

Embodiment 2

[0051] This example is used to prepare a low-serum, protein-free cell culture medium suitable for Vero cell growth.

[0052] 1. The formulation of the culture medium

[0053] Table 2 medium formula

[0054]

[0055]

[0056]

[0057] 2. Preparation method

[0058] (1) Get the dried components in the formula and put them into the ball mill tank respectively, and ball mill for 1.5 hours to get the culture medium dry powder;

[0059] (2) Dissolve the dry medium powder prepared in step (1) in 900ml ultrapure water, then dilute it to 1L with ultrapure water, and filter and sterilize it with a 0.22 μm filter membrane to obtain a Vero Low-serum protein-free liquid medium for cell growth, refrigerated at 8°C for later use.

Embodiment 3

[0061] This example is used to prepare a low-serum, protein-free cell culture medium suitable for Vero cell growth.

[0062] 1. The formulation of the culture medium

[0063] Table 3 medium formula

[0064]

[0065]

[0066]

[0067] 2. Preparation method

[0068] (1) Get the dried components in the formula and put them into the ball mill tank respectively, and ball mill for 3 hours to get the culture medium dry powder;

[0069] (2) Dissolve the dry medium powder prepared in step (1) in 900ml ultrapure water, then dilute it to 1L with ultrapure water, and filter and sterilize it with a 0.22 μm filter membrane to obtain a Vero Low-serum protein-free liquid medium for cell growth, refrigerated at 4°C for later use.

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Abstract

The invention belongs to the field of cell culture and particularly relates to a low-serum protein-free culture medium applicable to Vero cell growth and a preparation method thereof. The culture medium comprises amino acid, vitamin, inorganic salt, micro elements and other ingredients. The low-serum protein-free culture medium applicable to Vero cell growth and the preparation method of the low-serum protein-free culture medium have the advantages of being free of protein and animal source ingredients and definite in chemical ingredient, the interference to cell culture of serum is industrially reduced, the production cost is lowered, and great convenience is brought to industrial production of cells.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a low-serum protein-free medium suitable for Vero cell growth and a preparation method thereof. Background technique [0002] Vero cells are African green monkey kidney cells, which are aneuploid cells derived from rhesus monkey kidney cells. This cell line was isolated and cultured from kidney epithelial cells of African green monkeys by scholars Y. Yasumura and Y. Kawakita of Chiba University in Japan in 1962. Vero cell is a cell line for vaccine production approved by the World Health Organization and my country's biological products regulations. It has convenient source, high biological safety, can be continuously passaged in vitro, is sensitive to various viruses and has high virus proliferation titer, and has been approved by the World Health Organization. Recommended as an ideal cell-cell matrix for the production of human vaccines. [0003] The production process ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 陈文均徐念沁陈巧云
Owner 南京三生生物技术股份有限公司
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