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Serum-free medium for Madin-Darby canine kidney (MDCK) cell full suspension culture

A serum-free medium, full suspension technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc.

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

At present, there is no commercial MDCK cell serum-free medium in China

Method used

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  • Serum-free medium for Madin-Darby canine kidney (MDCK) cell full suspension culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0152] A serum-free medium for full suspension culture of MDCK cells. The raw materials are classified and dissolved according to their respective dissolution characteristics, 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 value of the resulting mixed solution is adjusted to 7.22. After constant volume, the serum-free medium for the full suspension culture of MDCK cells is obtained, wherein the final concentration of each raw material is:

[0153] Amino acid part:

[0154] L-alanine 100 mg / L,

[0155] L-arginine hydrochloride 120 mg / L,

[0156] L-Asparagine 5 mg / L,

[0157] L-Aspartic Acid 65 mg / L,

[0158] L-cysteine ​​monohydrate 10 mg / L,

[0159] L-cystine hydrochloride 25 mg / L,

[0160] L-glutamic acid 45 mg / L,

[0161] L-Glutamine 300 mg / L,

[0162] Glycine 70 mg / L,

[0163] L-histidine hydrochloride 110 mg / L,

[0164] L-Isoleucine 142 mg / L,

[0165] L-leucine 150 mg / L,

[016...

Embodiment 2

[0223] A serum-free medium for full suspension culture of MDCK cells. The raw materials are classified and dissolved according to their respective dissolution characteristics, and then the resulting solutions are mixed at 25.1° C., so that the final concentration of each raw material is as above, and the pH value of the resulting mixed solution is adjusted to 7.12. After constant volume, the serum-free medium for MDCK cell suspension culture is obtained, wherein the final concentration of each raw material is:

[0224] Amino acid part:

[0225] L-alanine 120 mg / L,

[0226] L-arginine hydrochloride 200 mg / L,

[0227] L-Asparagine 10 mg / L,

[0228] L-Aspartic Acid 80 mg / L,

[0229] L-cysteine ​​monohydrate 45 mg / L,

[0230] L-cystine hydrochloride 50 mg / L,

[0231] L-glutamic acid 72 mg / L,

[0232] L-Glutamine 405 mg / L,

[0233] Glycine 85 mg / L,

[0234] L-histidine hydrochloride 155 mg / L,

[0235] L-Isoleucine 180 mg / L,

[0236] L-leucine 180 mg / L,

[0237] L-Lysine hy...

Embodiment 3

[0294] A serum-free medium for full suspension culture of MDCK cells. The raw materials are mixed and ground into fine powder, and then the obtained fine powder is dissolved in distilled water at 28.6° C., so that the final concentration of each raw material is as above, and the obtained mixed solution is adjusted. When the pH value reaches 7.13, the serum-free medium for the full suspension culture of MDCK cells is obtained after constant volume, wherein the final concentration of each raw material is:

[0295] Amino acid part:

[0296] L-alanine 115.5 mg / L,

[0297] L-arginine hydrochloride 147.5 mg / L,

[0298] L-Asparagine 7.5 mg / L,

[0299] L-Aspartic Acid 75.65 mg / L,

[0300] L-cysteine ​​monohydrate 17.56 mg / L,

[0301] L-cystine hydrochloride 31.29 mg / L,

[0302] L-glutamic acid 50.35 mg / L,

[0303] L-Glutamine 365 mg / L,

[0304] Glycine 78.75 mg / L,

[0305] L-histidine hydrochloride 131.48 mg / L,

[0306] L-isoleucine 154.47 mg / L,

[0307] L-leucine 159.05 mg / L,...

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Abstract

The invention discloses a serum-free medium for Madin-Darby canine kidney (MDCK) cell full suspension culture. The serum-free medium for MDCK cell full suspension culture comprises an amino acid part, a vitamin part, an inorganic salt part and other additive parts. The serum-free medium has the beneficial effects that the serum-free medium for MDCK cell full suspension culture, provided by the invention, is high in cell culture density and clear in composition and does not contain animal serum, a downstream product is purified, the product quality is improved, and the serum-free medium is convenient to prepare and use and suitable for large-scale production of influenza vaccines and avian influenza vaccines.

Description

technical field [0001] The invention relates to a serum-free medium, in particular to a serum-free medium for full suspension culture of MDCK cells. Background technique [0002] MDCK cells (Madin-Darby canine kidney cells, MDCK) were isolated and cultured by Madin and Darby in 1958 from the kidney tissue of American Cocker Spaniel bitches. MDCK cells are currently widely used in the amplification and purification of a variety of viruses, such as reovirus, adenovirus, canine parvovirus, feline granulocytosis virus, and avian influenza virus. Due to their high virus infection efficiency and rapid proliferation, And it is not easy to mutate. MDCK cell line is recognized as one of the three cell lines most suitable for the production of influenza A and B influenza virus vaccines. [0003] At present, influenza vaccines in my country (including human influenza vaccines and animal influenza vaccines) are produced by the chicken embryo method. This process has defects such as lon...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 乔自林马忠仁令世鑫冯玉萍冯若飞李明生马祺
Owner 无锡市赛尔百灵生物技术有限公司
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