Cell culture medium and preparation method thereof

A culture medium and cell technology, applied in cell culture medium, cell culture active agents, biochemical equipment and methods, etc., can solve the problems of difficult separation, purification and detection of culture products, limited sources of animal serum, and restrictions on large-scale use, etc. Achieve the effect of improving cell proliferation, promoting wound healing, and good biocompatibility

Inactive Publication Date: 2018-08-21
广州瑞贝斯药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition of animal serum is complex, various biological size molecules are mixed together, and some components have not yet been clarified
Serum is very effective for cell growth, but it will cause some difficulties in the separation, purification and detection of cultured products in the later stage
In addition, the sources of high-quality animal serum are limited and the cost is high, which limits its large-scale use

Method used

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  • Cell culture medium and preparation method thereof
  • Cell culture medium and preparation method thereof
  • Cell culture medium and preparation method thereof

Examples

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preparation example Construction

[0025] Preparation of sericin solution: Sericin with a purity of more than 92% is dissolved in water or PBS buffer solution to obtain a sericin stock solution with a concentration greater than 100 mg / ml. The stock solution is used after being sterilized by conventional high temperature sterilization method or sterilized by 0.2 μm filter.

[0026] The phytohemagglutinin is selected from kidney bean phytohemagglutinin.

[0027] The preparation method of this culture medium comprises the following steps:

[0028] 4) Accurately weigh the dry powder of basic medium, chitosan quaternary ammonium salt, vitamin C, vitamin E, human serum albumin, and phytohemagglutinin of corresponding concentrations and dissolve them in an ultra-pure medium with a volume of 800ml and a temperature of 20-30°C. In water, stir until completely dissolved to obtain solution A;

[0029] 5) Add corresponding concentrations of sericin solution, epidermal growth factor, insulin-like growth factor, and L-glut...

Embodiment 1

[0033] The serum-free cell culture medium of this embodiment mainly includes the following components, DMEM / F12 medium 80v / v%, sericin solution 50mg / ml, chitosan quaternary ammonium salt solution 2mg / ml, human blood white Protein 35mg / ml, phytohemagglutinin 30mg / ml, vitamin C 20ug / ml, vitamin E 40ug / ml, epidermal growth factor 1.0ng / ml, insulin-like growth factor 5ng / ml, L-glutamine 3mmol / ml.

[0034] The preparation method of the culture medium in this embodiment is as described above and will not be described here.

Embodiment 2

[0036] The serum-free cell culture medium of the present invention mainly includes the following components: DMEM / F12 medium 85v / v%, sericin solution 25mg / ml, chitosan quaternary ammonium salt solution 2.5mg / ml, human blood white Protein 50mg / ml, phytohemagglutinin 20mg / ml, vitamin C 50ug / ml, vitamin E 50ug / ml, epidermal growth factor 0.5ng / ml, insulin-like growth factor 10ng / ml, L-glutamine 1mmol / ml.

[0037] The preparation method of the culture medium in this example is the same as that in Example 1, and will not be described here.

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Abstract

The invention discloses a cell culture medium. The cell culture medium is prepared from components as follows: 80v / v%-90v / v% of a DMEM / F12 culture medium, 25-500 mg / ml of a sericin solution, 2.0-5 mg / ml of a chitosan quaternary ammonium salt solution, 35-70 mg / ml of human serum albumin, 20-80 mg / ml of phytohemagglutinin, 20-100 mu g / ml of vitamin C, 20-80 mu g / ml of vitamin E, 0.5-10 ng / ml of an epidermal growth factor, 5-20 ng / ml of an insulin-like growth factor and 1-5 mmol / ml of L-glutamine. Cell growth can be promoted by use of sericin, and the chitosan quaternary ammonium salt solution isused for the cell culture medium, can improve the cell proliferation capability and tumor killing rate and can improve the cell growth capability when matched with sericin for use. Kidney bean lectinhas the functions of agglutinating blood cells, lymphocyte, spermatid and the like, and in in-vitro cell culture, phytolectin can proliferate lymphocyte; in the whole experiment, the phytolectin canimprove the immune capability of a body; under the synergistic action of sericin, the chitosan quaternary ammonium salt solution and lectin, the proliferation rate, the survival rate and the tumor killing rate of T lymphocyte are remarkably increased.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cell culture medium and a preparation method thereof. Background technique [0002] Cell culture medium is not only the basic material for supplying cell nutrition and promoting cell reproduction and proliferation in cultured cells, but also the living environment for the growth and reproduction of cultured cells. [0003] In 1951, Earle developed a synthetic medium (MEM) for the growth of animal cells in vitro. There are quite a few types of synthetic media. The composition of the synthetic medium is known, which facilitates the control of the experimental conditions. However, compared with the natural medium, some natural unknown components cannot be replaced by known chemical components. Therefore, the basic synthetic medium used in cell culture must also add a certain amount of natural medium components to overcome the synthetic culture. Insufficient base. The most...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/0037C12N2500/32C12N2500/34C12N2500/38C12N2501/105C12N2501/11C12N2501/59C12N2501/998
Inventor 艾譞林杰庚
Owner 广州瑞贝斯药业有限公司
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