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Chinese hamster ovary culture medium as well as preparation method and application thereof

A technology of ovarian cells and Chinese hamsters, applied in biochemical equipment and methods, tissue culture, microorganisms, etc., can solve the problems of cultured cell pollution threats, uncertain chemical composition, and limited cell research and development

Inactive Publication Date: 2011-04-20
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the uncertain chemical composition of the existing Chinese hamster ovary cell culture medium, the use of animal source components or protein, so that the cultured cells have potential pollution threats, and the cost is high, increasing the separation and purification Difficulty, to a certain extent limit the development of cell research and other defects, provide a Chinese hamster ovary cell culture medium and its preparation method and its preparation method and application

Method used

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  • Chinese hamster ovary culture medium as well as preparation method and application thereof

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Embodiment 1

[0021] Prepare the animal-derived component-free, protein-free, and chemically defined culture medium of the present invention according to the following component concentrations, and dissolve each component in water, wherein ferric citrate needs to be heated and dissolved in a boiling water bath before being uniformly mixed with other substances; Caprylic acid is dissolved in absolute ethanol and then mixed uniformly; after uniform mixing, a 0.22 micron nitrocellulose membrane is used for filter sterilization.

[0022] The Chinese hamster ovary cell culture medium of the present invention contains: 72mg / L L-tryptophan, 50mg / L L-cystine, 87.6mg / L L-cysteine, 130.2mg / L L-tyrosine Sodium salt dihydrate, 324.6mg / LL-isoleucine, 376.8mg / L L-leucine, 190.6mg / L L-valine, 321.2mg / L L-arginine, 206.1mg / L L -Histidine, 425.3mg / L L-lysine, 84.7mg / L L-methionine, 196.7mg / L L-phenylalanine, 182.6mg / L L-glutamic acid, 213mg / L L-day Paragine, 13mg / L glycine, 210mg / L L-proline, 7mg / L L-alani...

Embodiment 2

[0026] On the basis of the ingredients contained in the culture medium in Example 1, add 1.5mg / L stannous chloride dihydrate, 0.08mg / L ammonium vitriol, 0.02mg / L barium acetate, 0.01mg / L silver nitrate, 0.01mg / L sodium silicate, 0.7mg / L aluminum chloride hexahydrate, 0.003mg / L cadmium sulfate octahydrate, 0.1mg / L cobalt chloride hexahydrate, 0.2mg / L manganese sulfate monohydrate, 0.033mg / L tetrahydrate Ammonium molybdate, 0.45mg / L nickel chloride hexahydrate, 0.00002mg / L potassium bromide, 0.0058mg / L sodium fluoride, 0.00023mg / L potassium iodide, 0.0035mg / L arachidonic acid, 0.52mg / L cholesterol . Among them, ferric citrate and ammonium vanitate need to be heated and dissolved in a boiling water bath before being evenly mixed with other substances; lipoic acid and stannous chloride are dissolved in absolute ethanol, then evenly mixed, and then 0.22 micron nitrocellulose membrane is used Perform filter sterilization.

[0027] CHO cells were inserted into the prepared medium ...

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Abstract

The invention discloses a Chinese hamster ovary culture medium as well as a preparation method and application thereof. The culture medium comprises ascorbic acid, pyridoxine hydrochloride, vitamin B12, nicotinamide, riboflavin, thiamine chloride, choline chloride, folic acid, calcium pantothenate, sodium pyruvate, reduced glutathione, inositol, biotin, hypoxanthine, putrescine, lipoic acid, linoleic acid, thymine, glucose, HEPES (2-hydroxyethyl) buffer solution, dextran sulphate, Pluronic-F68, various amino acids and salts thereof and various inorganic salts. The culture medium has low cost, and the component does not contain animal origin ingredients, does not contain protein and has definite chemical ingredients. All ingredients are cooperated and blended, and the Chinese hamster ovary culture medium can satisfy the basic growth requirement of cells, and can effectively regulate, transfer and control the culture of cells so as to realize good high-density cell culture. Compared with the culture effect of the existing culture medium, the culture effect of culture medium disclosed in the invention is at least equivalent to even superior to the existing culture medium.

Description

technical field [0001] The invention relates to a Chinese hamster ovary cell culture medium and its preparation method and application. Background technique [0002] Chinese hamster ovary cells (CHO cells) are currently the most widely used mammalian cell line and have been used to produce various genetically engineered protein products, for example, tissue plasminogen activator (t-PA) , human erythropoietin (rHuEPO), hepatitis B surface antigen (HBsAg), human thrombopoietin (rhTPO), various recombinant antibodies, etc. [0003] CHO cells are usually cultured in a synthetic medium supplemented with fetal calf serum or calf serum. However, the added serum is a substance with unclear composition, which contains many proteins and other macromolecular substances with uncertain composition, which affects the separation and purification of the product and reduces the yield and purity of the product. At the same time, there is a potential risk of virus infection in the serum deri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06
Inventor 易小萍张大鹤孙祥明张元兴
Owner EAST CHINA UNIV OF SCI & TECH
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