Method for preparing serum-free soybean protein peptide animal cell medium

A soybean protein peptide and animal cell technology, applied in the field of preparation of serum-free or low-serum medium, can solve problems such as lethal damage, and achieve the effects of simplifying the separation and purification process, improving product purity, and reducing impurity content

Inactive Publication Date: 2010-12-15
乐能生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the existing serum-free medium cannot directly cultivate cells with poor growth conditions. Most cells must gradually reduce the serum concentration to adapt to the existing serum-free medium and are easily affected by external factors, which can easily cause lethality. damage problem, while providing a serum-free soy protein peptide animal cell culture medium

Method used

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  • Method for preparing serum-free soybean protein peptide animal cell medium
  • Method for preparing serum-free soybean protein peptide animal cell medium

Examples

Experimental program
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example 1

[0011] Calcium chloride 100mg, potassium chloride 100mg, anhydrous magnesium sulfate 50mg, sodium chloride 3000mg, anhydrous sodium dihydrogen phosphate 100mg, L-glutamine 100mg, D-glucose 500mg, phenol red 5mg, D-calcium pantothenate 0.50mg, choline chloride 0.50mg, folic acid 0.5mg, i-inositol 1mg, nicotinamide 0.5mg, pyridoxal hydrochloride 0.5mg, riboflavin 0.05mg, sulfur hydrochloride 0.5mg and soybean protein peptide 2000mg with distilled water The molecular weight of the soybean protein peptide is less than or equal to 3000 Daltons, and the soybean protein peptide contains 18 amino acids or more.

[0012] This medium can be used to culture CHO cells, NSO cells, Sp2 / O cells, hybridoma cells, HEK293 cells, MDCK cells, HeLa cells, MDBK cells, etc.

[0013] For special cells that must be cultured with serum, such as Marcl45 cells, BHK21, VERO cells, bone marrow mesenchymal stem cells, etc., serum with a mass fraction of less than 3% is added to the serum-free soy protein pe...

example 2

[0017] Calcium chloride 400mg, potassium chloride 500mg, anhydrous magnesium sulfate 150mg, sodium chloride 8000mg, anhydrous sodium dihydrogen phosphate 200mg, L-glutamine 500mg, D-glucose 2000mg, phenol red 20mg, D-calcium pantothenate 5mg, choline chloride 5mg, folic acid 5mg, i-inositol 5mg, nicotinamide 5mg, pyridoxal hydrochloride 5mg, riboflavin 1mg, sulfur hydrochloride 5mg and soybean protein peptide 8000mg with distilled water to make up to 1000mL; The molecular weight of soybean protein peptide is less than or equal to 3000 Daltons, and soybean protein peptide contains 18 kinds of amino acids or more.

[0018] This medium can be used to culture CHO cells, NSO cells, Sp2 / O cells, hybridoma cells, HEK293 cells, MDCK cells, HeLa cells, MDBK cells, etc.

[0019] For special cells that must be cultured with serum, such as Marcl45 cells, BHK21, VERO cells, bone marrow mesenchymal stem cells, etc., serum with a mass fraction of less than 3% is added to the serum-free soy p...

example 3

[0021] In this embodiment, HaLe cells (African female cervical cancer cells) are cultured with serum-free soy protein peptide animal cell culture medium.

[0022] The culture medium of Example 1 and MEM cell culture medium (product code: MD600, batch number: 070512) were used to directly culture the recovered HaLe cells at 37°C, and the morphology, growth and proliferation of the cells in each group were observed with a microscope.

[0023] Table 1

[0024]

[0025]

[0026] The percentage of cell division cultured in serum-free soy protein animal cell medium is significantly better than that of ordinary MEM cell medium, and the time for cells to grow into monolayers in serum-free soy protein peptide animal cell medium is the same as that in traditional medium, and the cells The shape is basically the same.

[0027] The nutrient composition of serum-free soy protein peptide animal cell medium is better than that of ordinary serum medium. The cells in serum-free soy prot...

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Abstract

The invention discloses a method for preparing a serum-free soybean protein peptide animal cell medium, and relates to a serum-free or low-serum collective cell medium. The serum-free soybean protein peptide animal cell medium is prepared from calcium chloride, potassium chloride, anhydrous magnesium sulfate, sodium chloride, anhydrous sodium dihydrogen phosphate, L-glutamine, D-glucose, phenol red, D-calcium pantothenate, choline chloride, folic acid, i-inositol, niacinamide, pyridoxal hydrochloride, lactoflavin, thiamine hydrochloride, soybean protein peptide and distilled water. The method of the invention solves the problem that the conventional serum-free medium cannot be directly used for cultivating cells in the poor growth condition, and a large number of the cells can adapt to the conventional serum-free medium only when the serum concentrations thereof are gradually reduced, and the cells are easily affected by external factors, so fatal damages are easily caused. The method of the invention has the advantages that the cells can directly be cultivated in the medium of the invention without gradual domestication and cultivation, so the culture period of the cell is shortened and the interference of other factors is avoided; meanwhile, the virus pollution brought by serums is reduced, and the survival rate of the cells is over 94 percent.

Description

1. Technical field [0001] The present invention relates to the preparation of serum-free or low-serum culture medium, in particular to a preparation method (virus host) of a serum-free soybean protein peptide animal cell culture medium. 2. Background technology [0002] Animal cells are expression vectors for viruses and proteins, and the quality of cells directly affects the amount of expression. Only a good medium can screen and domesticate high-quality cells. Ordinary cell culture medium contains calf serum. Since the serum is produced in batches, the difference between batches is very large and the quality is unstable. Moreover, the serum contains some substances that are toxic to cells, which will affect the growth of cells and even cause Cell death also increases the difficulty of separation and purification in production, and is especially unsuitable for the study of the effect of growth factors on cells, the study of the function of cell secretions, and the preparati...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 王东典梁金钟王淑杰
Owner 乐能生物工程股份有限公司
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