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A low-serum cell culture medium with wide adaptability and preparation method thereof

An adaptable, low-serum technology, applied in the field of biochemistry, can solve the problems of rising animal serum prices and aggravating the economic burden of enterprises, and achieve the effects of not reducing cell viability, small serum dosage, and wide adaptability.

Active Publication Date: 2020-08-25
山东巨山能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional cell culture has a great demand for animal serum, and due to changes in the animal husbandry industry, the price of animal serum continues to rise, increasing the economic burden on enterprises

Method used

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  • A low-serum cell culture medium with wide adaptability and preparation method thereof
  • A low-serum cell culture medium with wide adaptability and preparation method thereof
  • A low-serum cell culture medium with wide adaptability and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The steps of the present invention's low-serum cell culture medium are as follows:

[0033] (1) Preparation solution: 0.0035 parts by mass of biotin, 0.2 parts by mass of riboflavin, 0.7 parts by mass of vitamin B12, 0.0012 parts by mass of copper sulfate pentahydrate, 0.05 parts by mass of ferric nitrate nonahydrate, 0.5 parts by mass of ferrous sulfate heptahydrate , 0.75 parts by mass of zinc sulfate heptahydrate, 0.0003 parts by mass of ammonium metavanadate, 0.00006 parts by mass of manganous chloride, 0.003 parts by mass of sodium selenite, 2 parts by mass of ethanolamine, 1 part by mass of linoleic acid, 0.08 parts by mass of lipoic acid , 0.06 parts by mass of putrescine dihydrochloride and 0.3 parts by mass of thymidine are dissolved in 1000 parts by mass of ultrapure water to prepare a solution;

[0034] (2) drying: the solution prepared in step (1) is mixed with 3000 parts by mass of D-glucose and then dried;

[0035] (3) Ball milling: take materials and mix...

Embodiment 2

[0040] The difference between this embodiment and Example 1 is: in step (1), 0.002 mass parts biotin, 0.1 mass parts riboflavin, 0.2 mass parts vitamin B12, 0.0005 mass parts copper sulfate pentahydrate, 0.02 mass parts nine Ferric nitrate water, 0.2 mass parts ferrous sulfate heptahydrate, 0.5 mass parts zinc sulfate heptahydrate, 0.0001 mass parts ammonium metavanadate, 0.00002 mass parts manganous chloride, 0.002 mass parts sodium selenite, 1 mass part ethanolamine, 3 parts by mass of linoleic acid, 0.05 parts by mass of lipoic acid, 0.05 parts by mass of putrescine dihydrochloride and 0.1 part by mass of thymidine are dissolved in 940 parts by mass of ultrapure water to prepare a solution;

[0041] In step (2), the solution prepared in step (1) is mixed with 1000 parts by mass of D-glucose and then dried;

[0042] In step (3), take material and mix by following mass parts:

[0043]

[0044]

[0045] Then mix with the mixture dried in step (2) by a three-dimensional ...

Embodiment 3

[0047] The difference between this embodiment and Example 1 is: in step (1), 0.004 mass parts biotin, 0.3 mass parts riboflavin, 0.8 mass parts vitamin B12, 0.0015 mass parts copper sulfate pentahydrate, 0.06 mass parts nine Iron nitrate water, 0.6 parts by mass of ferrous sulfate heptahydrate, 1 part by mass of zinc sulfate heptahydrate, 0.0005 parts by mass of ammonium metavanadate, 0.0001 parts by mass of manganese chloride, 0.007 parts by mass of sodium selenite, 1.5 parts by mass of ethanolamine, 5 parts by mass of linoleic acid, 0.12 parts by mass of lipoic acid, 0.1 part by mass of putrescine dihydrochloride and 0.5 parts by mass of thymidine are dissolved in 1060 parts by mass of ultrapure water to prepare a solution;

[0048] In step (2), the solution prepared in step (1) is mixed with 4500 parts by mass of D-glucose and then dried;

[0049] In step (3), take material and mix by following mass parts:

[0050]

[0051]

[0052] Then mix with the mixture dried in...

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Abstract

The invention discloses a low-serum cell culture medium with universal adaptability and a preparation method thereof, and belongs to the technical field of biochemistry, the preparation method comprises the following steps: (1), compounding solvent: dissolving trace materials into 940 to 1060 parts by mass of ultrapure water, compounding to be a solvent; (2), drying: mixing the solvent obtained in the step (1) with 1000 to 4500 parts by mass of D-glucose and drying; (3), ball milling: taking other raw materials and mixing uniformly with dried mixed materials in the step (2), then performing ball-milling, sieving through a 100-mesh screen, taking the sieved parts to prepare the culture medium. The culture medium has universal adaptability, can be applied to various cell lines such as VERO, BHK-21, A549, ST, PK-15; transition is not needed when cells is transferred from a high-serum culture medium to the low-serum cell culture medium provided by the invention during a culture process, the cells can adapt quickly, and the operation is simple and convenient.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to a low-serum cell culture medium with wide adaptability and a preparation method thereof. Background technique [0002] Cell culture has been widely used in basic and applied research in biology and medicine, and has also achieved large-scale production and application in the biopharmaceutical industry. It is one of the most important basic sciences. [0003] With the rapid development of biomedicine, the demand for vaccines, monoclonal antibodies and other biomedical products in the market continues to increase, and more and more biopharmaceutical companies have emerged as the times require. Traditional cell culture has a great demand for animal serum, and due to changes in the animal husbandry industry, the price of animal serum continues to rise, which increases the economic burden on enterprises. [0004] The low-serum cell culture medium not only contains a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00
CPCC12N5/0018C12N2500/05C12N2500/25C12N2500/30C12N2500/32C12N2500/34C12N2500/35C12N2500/38C12N2500/40C12N2500/46
Inventor 齐智明恒磊孟双有成芳硕孟丹丹信健房圆瑗牛欣桐朱宁
Owner 山东巨山能源科技有限公司
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