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Serum-free medium for full suspension cultivation of BHK (Baby Hamster Kidney)-21 cell

A technology of BHK-21 and serum-free medium, which is applied in tissue culture, animal cells, vertebrate cells, etc., and can solve the problems of unclear composition of medium and high price

Active Publication Date: 2014-02-05
无锡市赛尔百灵生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, the formula of foreign cell serum-free medium is confidential and the price is high, and the ingredients of the existing medium (domestic and foreign) on the market are not clear

Method used

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  • Serum-free medium for full suspension cultivation of BHK (Baby Hamster Kidney)-21 cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0127] A serum-free medium for full suspension culture of BHK-21 cells. The raw materials are classified and dissolved according to their respective dissolution characteristics, and then the resulting solutions are mixed at 23.1° C., so that the final concentrations of the raw materials are as above, and the pH of the resulting mixture is adjusted. value to 7.15, the serum-free medium for BHK-21 cell full suspension culture can be obtained after constant volume, wherein, the final concentration of each raw material is:

[0128] Amino acid part:

[0129] L-alanine 200 mg / L,

[0130] L-arginine hydrochloride 350 mg / L,

[0131] L-Asparagine 200 mg / L,

[0132] L-Aspartic Acid 50 mg / L,

[0133] L-cysteine ​​monohydrate 10 mg / L,

[0134] L-cystine hydrochloride 25 mg / L,

[0135] L-glutamic acid 100 mg / L,

[0136] L-Glutamine 300 mg / L,

[0137] Glycine 150 mg / L,

[0138] L-histidine hydrochloride 155 mg / L,

[0139] L-Isoleucine 100 mg / L,

[0140] L-leucine 150 mg / L,

[014...

Embodiment 2

[0187] A serum-free medium for full suspension culture of BHK-21 cells. The raw materials are classified and dissolved according to their respective dissolution properties, and then the resulting solutions are mixed at 25.9° C., so that the final concentration of each raw material is as above, and the pH of the resulting mixture is adjusted. value to 7.21, the serum-free medium for BHK-21 cell full suspension culture can be obtained after constant volume, wherein, the final concentration of each raw material is:

[0188] Amino acid part:

[0189] L-alanine 300 mg / L,

[0190] L-arginine hydrochloride 500 mg / L,

[0191] L-Asparagine 300 mg / L,

[0192] L-Aspartic Acid 85 mg / L,

[0193] L-cysteine ​​monohydrate 45 mg / L,

[0194] L-cystine hydrochloride 50 mg / L,

[0195] L-glutamic acid 155 mg / L,

[0196] L-Glutamine 405 mg / L,

[0197] Glycine 200 mg / L,

[0198] L-histidine hydrochloride 200 mg / L,

[0199] L-Isoleucine 150 mg / L,

[0200] L-leucine 180 mg / L,

[0201] L-Ly...

Embodiment 3

[0247] A serum-free medium for full suspension culture of BHK-21 cells. The raw materials are mixed and ground into a fine powder, and then the obtained fine powder is dissolved in distilled water at 25.3° C., so that the final concentration of each raw material is as above, and the resulting mixture is adjusted. The pH value of the solution is 7.23, and after constant volume, the serum-free medium for the whole suspension culture of BHK-21 cells is obtained, wherein the final concentration of each raw material is:

[0248] Amino acid part:

[0249] L-alanine 213.1 mg / L,

[0250] L-arginine hydrochloride 372.75 mg / L,

[0251] L-Asparagine 153.5 mg / L,

[0252] L-Aspartic Acid 75.65 mg / L,

[0253] L-cysteine ​​monohydrate 22.71 mg / L,

[0254] L-cystine hydrochloride 31.29 mg / L,

[0255] L-glutamic acid 109.4 mg / L,

[0256] L-Glutamine 365 mg / L,

[0257] Glycine 178.75 mg / L,

[0258] L-histidine hydrochloride 177.64 mg / L,

[0259] L-Isoleucine 134 mg / L,

[0260] L-leuci...

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Abstract

The invention discloses a serum-free medium for full suspension cultivation of a BHK (Baby Hamster Kidney)-21 cell. The serum-free medium for full suspension cultivation of the BHK-21 cell comprises an amino acid part, a vitamin part, an inorganic salt part and a part of other additives. The medium disclosed by the invention has the beneficial effects that the serum-free medium for full suspension cultivation of the BHK-21 cell is high in culture cell density, clear in component and free from animal serum, facilitates purification of downstream products, and improves the product quality. The medium is convenient to prepare and use, and is applicable to production of biological products on a large scale.

Description

technical field [0001] The invention relates to a serum-free medium, in particular to a serum-free medium for BHK-21 cell suspension culture. Background technique [0002] BHK-21 cells are Syrian hamster kidney cells, English name: Baby Hamster Kidney cell referred to as BHK-21 or BHK21, the cell line was established in 1961 by Macpherson IA, Stoker MGP, derived from 5 1-day-old unidentified sex Hamster kidney, single-cell cloned in 1963. The original cell line is fibroblast, which grows in an anchorage-dependent manner, but after numerous passages and domestication, it can grow in suspension and be used for the production of viral vaccines, such as foot-and-mouth disease vaccine, rabies vaccine, and Japanese encephalitis vaccine. The use of serum-free culture technology will improve the quality of the vaccine, which is mainly reflected in the following aspects: 1) Animal serum is no longer used in the process (bovine serum is added to the culture medium when the vaccine is...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/02
Inventor 令世鑫马忠仁李明生冯若飞王家敏马祺冯玉萍乔自林
Owner 无锡市赛尔百灵生物技术有限公司
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