Serum-free animal origin-free culture medium for suspension culture of BHK-21 cells and preparation method of serum-free animal origin-free culture medium

A BHK-21, suspension culture technology, applied in the direction of animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of long doubling time, low density, poor medium performance, etc., achieve rapid cell growth and proliferation, promote Proliferation speed, high vitality effect

Active Publication Date: 2015-11-04
内蒙古金源康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] At present, the price of foreign serum-free medium products is high and the formula is confidential. At the same time, there is a lack of serum-free medium specifically for BHK cell culture at home and abroad, and the performance of the medium is poor. For example, the latest patent CN103555658A publis

Method used

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  • Serum-free animal origin-free culture medium for suspension culture of BHK-21 cells and preparation method of serum-free animal origin-free culture medium
  • Serum-free animal origin-free culture medium for suspension culture of BHK-21 cells and preparation method of serum-free animal origin-free culture medium
  • Serum-free animal origin-free culture medium for suspension culture of BHK-21 cells and preparation method of serum-free animal origin-free culture medium

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] This embodiment is a serum-free low-protein medium (JYK-SFM) suitable for BHK-21 cell suspension culture, which includes 22 kinds of amino acids, 12 kinds of inorganic salts, 12 kinds of vitamins, 5 kinds of lipids, 4 kinds of One protein hydrolyzate, 14 kinds of additives; all materials used were purchased from Sigma Company unless otherwise specified. The specific ingredients are as follows:

[0103] (1) Amino acids:

[0104] L-alanine 100mg / L,

[0105] L-Glutamine 200mg / L,

[0106] L-arginine hydrochloride 200mg / L,

[0107] L-aspartic acid 150mg / L,

[0108] L-Asparagine 450mg / L,

[0109] L-cysteine ​​monohydrate 10mg / L,

[0110] L-glutamic acid 50mg / L,

[0111] L-isoleucine 100mg / L,

[0112] L-leucine 130mg / L,

[0113] L-lysine hydrochloride 250mg / L,

[0114] L-methionine 90mg / L,

[0115] L-phenylalanine 60mg / L,

[0116] L-proline 100mg / L,

[0117] L-serine 100mg / L,

[0118] L-Threonine 250mg / L,

[0119] L-Tryptophan 100mg / L,

[0120] L-tyrosine disod...

Embodiment 2

[0179] This embodiment is a serum-free low-protein medium (JYK-SFM) suitable for BHK-21 cell suspension culture, which includes 22 kinds of amino acids, 12 kinds of inorganic salts, 12 kinds of vitamins, 5 kinds of lipids, 4 kinds of One protein hydrolyzate and 14 additives; all materials used were purchased from Sigma Company unless otherwise specified, and the specific ingredients are as follows:

[0180] (1) Amino acids:

[0181] L-alanine 150mg / L,

[0182] L-Glutamine 275mg / L,

[0183] L-arginine hydrochloride 260mg / L,

[0184] L-Aspartic Acid 100mg / L,

[0185] L-Asparagine 300mg / L,

[0186] L-cysteine ​​monohydrate 35mg / L,

[0187] L-glutamic acid 100mg / L,

[0188] L-isoleucine 180mg / L,

[0189] L-Leucine 150mg / L,

[0190] L-lysine hydrochloride 100mg / L,

[0191] L-methionine 50mg / L,

[0192] L-phenylalanine 35mg / L,

[0193] L-proline 80mg / L,

[0194] L-serine 200mg / L,

[0195] L-Threonine 150mg / L,

[0196] L-tryptophan 150mg / L,

[0197] L-tyrosine disodium...

Embodiment 3

[0256] This embodiment is a serum-free low-protein medium (JYK-SFM) suitable for BHK-21 cell suspension culture, which includes 22 kinds of amino acids, 12 kinds of inorganic salts, 12 kinds of vitamins, 5 kinds of lipids, 4 kinds of One protein hydrolyzate, 14 kinds of additives; all materials used were purchased from Sigma Company unless otherwise specified. The specific ingredients are as follows:

[0257] (1) Amino acids:

[0258] L-alanine 180mg / L,

[0259] L-Glutamine 300mg / L,

[0260] L-arginine hydrochloride 300mg / L,

[0261] L-aspartic acid 120mg / L,

[0262] L-Asparagine 400mg / L,

[0263] L-cysteine ​​monohydrate 50mg / L,

[0264] L-glutamic acid 150mg / L,

[0265] L-isoleucine 300mg / L,

[0266] L-Leucine 100mg / L,

[0267] L-lysine hydrochloride 350mg / L,

[0268] L-methionine 35mg / L,

[0269] L-phenylalanine 10mg / L,

[0270] L-proline 155mg / L,

[0271] L-serine 150mg / L,

[0272] L-Threonine 200mg / L,

[0273] L-tryptophan 130mg / L,

[0274] L-tyrosine disodi...

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Abstract

The invention discloses a serum-free animal origin-free culture medium for suspension culture of BHK-21 cells and a preparation method of the serum-free animal origin-free culture medium. According to the serum-free animal origin-free culture medium for suspension culture of the BHK-21 cells and the preparation method of the serum-free animal origin-free culture medium, through the addition and component optimization of amino acids, inorganic salt, vitamins, lipids, hydrolysates and the rest of additives in the culture medium, the multiplication speed of the BHK-21 cells is remarkably improved, the doubling time is shortened, and the high activity is maintained while the cells reach higher growth density. The culture medium is suitable for high-density suspension culture of the BHK-21 cells, low in cost, free of serum, free of animal origins and low in protein, the biological function of the serum is completely replaced through the addition of the ingredients including the lipids, the vitamins, the microelements, recombinant protein and the hydrolysates,, and some negative effects caused by the serum to later purification and separation of an antigen are avoided. Through optimization on the components of the culture medium, the multiplication speed of the BHK-21 cells in suspension culture is higher, the cell density is higher, the cell activity is higher, and the BHK-21 cells are more suitable for large-scale industrial production.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a serum-free and animal-source-free culture medium for suspension culture of BHK-21 cells and a preparation method thereof. Background technique [0002] The BHK-21 cell line is the Syrian hamster kidney cell, English name: Baby Hamster Kidney Cell, in 1961, the British I.A.Macpherson and M.G.P Stocker established the passage cells with the kidney of a 1-day-old Syrian hamster. The commonly used clone 13 is derived from 5 Kidney cells that are only one day old and whose gender characteristics are not particularly obvious, namely: BHK-21C13, the full Chinese name is: Syrian Hamster Kidney Cell Clone 13, hereinafter referred to as BHK-21 cells. BHK-21 cell is a cell line approved by the World Health Organization and China Biological Products Regulations for the production of vaccines, and can be used for the propagation and purification of various viruses, such as foot a...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 赵峻岭
Owner 内蒙古金源康生物工程股份有限公司
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