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Serum-free culture medium

A serum-free culture medium, culture medium technology, applied in cell culture medium, culture process, tissue culture and other directions, can solve the problems of low cell expansion, short survival time, low antibody purity, etc. The effect of maintaining cell viability and high expansion rate

Inactive Publication Date: 2018-11-30
BEIJING VBIOSCI INC +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the commercially available culture medium has low expansion factor, short survival time, low activity, low yield, and low antibody purity, which cannot meet the problems of clinical application.

Method used

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  • Serum-free culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A serum-free medium comprising the following concentrations of raw materials:

[0039] DMEM / F12;

[0040] and add in DMEM / F12:

[0041] Fibroblast growth factor 0.1ng / mL, recombinant human epidermal growth factor (EGF) 0.1ng / mL, nerve growth factor 0.1ng / mL, transforming growth factor 0.1ng / mL, leukemia inhibitory factor 0.1ng / mL, insulin-like Growth factor-10.1ng / mL, hydrocortisone 12μg / L, insulin 2μg / L, progesterone 1μg / L, dexamethasone 1μg / L, serum albumin 1mg / mL, transferrin 1mg / mL, fetuin Protein 1mg / mL, glutamine 0.01g / L, reduced glutathione 0.1g / L, non-essential amino acid 0.01g / L, linoleic acid 0.5mg / L, cholesterol 0.01mg / L, ethanolamine 0.5mg / L L, β-mercaptoethanol 0.5mg / L, Tween-80 5mg / L, manganese chloride 1μg / L, sodium stannate 1μg / L, ferric citrate 10μg / L, zinc sulfate 10μg / L, biotin 0.01 mg / L, folic acid 0.1mg / L, riboflavin 0.1mg / L, vitamin B 12 0.1mg / L, Vitamin C 0.1mg / L, Vitamin E 0.1mg / L.

Embodiment 2

[0043] A serum-free medium comprising the following concentrations of raw materials:

[0044]DMEM / F12;

[0045] and add in DMEM / F12:

[0046] Fibroblast growth factor 25ng / mL, recombinant human epidermal growth factor (EGF) 25ng / mL, nerve growth factor 25ng / mL, transforming growth factor 25ng / mL, leukemia inhibitory factor 25ng / mL, insulin-like growth factor-125ng / mL mL, hydrocortisone 30 μg / L, insulin 10 μg / L, progesterone 10 μg / L, dexamethasone 10 μg / L, serum albumin 25 mg / mL, transferrin 25 mg / mL, fetuin 25 mg / mL, gluten Aminoamide 0.05g / L, reduced glutathione 0.75g / L, non-essential amino acid 0.05g / L, linoleic acid 0.75mg / L, cholesterol 0.05mg / L, ethanolamine 0.75mg / L, β-mercaptoethanol 0.75mg / L, Tween-80 7.5mg / L, manganese chloride 5μg / L, sodium stannate 5μg / L, ferric citrate 50μg / L, zinc sulfate 50μg / L, biotin 0.1mg / L, folic acid 1.5mg / L, riboflavin 0.15mg / L, vitamin B 12 0.4mg / L, Vitamin C 0.4mg / L, Vitamin E 0.4mg / L.

Embodiment 3

[0048] A serum-free medium comprising the following concentrations of raw materials:

[0049] DMEM / F12;

[0050] and add in DMEM / F12:

[0051] Fibroblast growth factor 50ng / mL, recombinant human epidermal growth factor (EGF) 50ng / mL, nerve growth factor 50ng / mL, transforming growth factor 50ng / mL, leukemia inhibitory factor 50ng / mL, insulin-like growth factor-150ng / mL mL, hydrocortisone 60 μg / L, insulin 20 μg / L, progesterone 20 μg / L, dexamethasone 20 μg / L, serum albumin 50 mg / mL, transferrin 50 mg / mL, fetuin 50 mg / mL, gluten Aminoamide 0.1g / L, reduced glutathione 1.5g / L, non-essential amino acid 0.1g / L, linoleic acid 1mg / L, cholesterol 0.1mg / L, ethanolamine 1mg / L, β-mercaptoethanol 1mg / L L. Tween-80 10mg / L, Manganese Chloride 10μg / L, Sodium Stannate 10μg / L, Ferric Citrate 100μg / L, Zinc Sulfate 100μg / L, Biotin 0.2mg / L, Folic Acid 3mg / L, Riboflavin 0.3mg / L, vitamin B 12 0.8mg / L, Vitamin C 0.8mg / L, Vitamin E 0.8mg / L.

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Abstract

The invention discloses a serum-free culture medium which comprises the following components of DMEM / F12 and growth factors of 0.6-300 ng / mL, hormone of 15-120 [mu]g / L, protein of 3-150 mg / mL, amino acid and peptide of 0.1-1.7 g / L, lipid compound of 6.51-13.1mg / L, trace elements of 22-220 [mu]g / L, and vitamin of 0.5-5.9 mg / L, wherein the growth factors of 0.6-300 ng / mL, the hormone of 15-120 [mu]g / L, the protein of 3-150 mg / mL, the amino acid and peptide of 0.1-1.7 g / L, the lipid compound of 6.51-13.1mg / L, the trace elements of 22-220 [mu]g / L, and the vitamin of 0.5-5.9 mg / L are added in the DMEM / F12. The invention relates to a cell culture medium, in particular to a culture medium for culturing suspension cells, the culture medium for the culturing suspension cells is used for the expression of recombinant proteins, the culture medium of the invention does not contain serum components at all, and the purity of the produced antibodies is greatly improved; meanwhile, the culture mediumhas high amplification rate and high antibody expression capacity; and the culture medium can maintain cell viability for a long time, and the cell culture time is greatly prolonged so that the production time can be greatly prolonged, and the antibody production is improved.

Description

technical field [0001] The invention relates to a serum-free culture medium for cells, which belongs to the technical field of cell culture. Background technique [0002] Serum-free medium refers to a synthetic medium that can maintain the growth and reproduction of cells in vitro for a long time without adding serum. It can be used to cultivate mammalian cells, such as Chinese hamster ovary cells (CHO), canine kidney cells (MDCK), Monkey kidney cells (Vero), baby hamster kidney cells (BHK), etc. [0003] At present, cell culture has been widely used to produce a variety of biologically active products, such as virus vaccines, monoclonal antibodies, non-antibody immunomodulators, polypeptide growth factors, hormones, enzymes, tumor-specific antigens, etc. These products can be produced by normal or transformed and genetically engineered cells. [0004] Traditionally, media used for cell culture required the addition of serum as a universal nutrient required for growth and ...

Claims

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Application Information

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IPC IPC(8): C12N5/00
CPCC12N5/0037C12N2500/12C12N2500/20C12N2500/22C12N2500/24C12N2500/25C12N2500/32C12N2500/36C12N2500/38C12N2500/44C12N2500/46C12N2501/105C12N2501/11C12N2501/115C12N2501/13C12N2501/15C12N2501/235C12N2501/392C12N2501/855C12N2501/998C12N2501/999
Inventor 侯野张乾顺邵瑞华黄广达王璐赵英杰韩丹张志豪宋春雨
Owner BEIJING VBIOSCI INC
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