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Epidermal stem cell amplification culture medium as well as preparation method and application thereof

A technology of epidermal stem cells and medium, applied in the field of stem cells, can solve the problems of fast cell differentiation, limited cell number, poor stability, etc.

Inactive Publication Date: 2021-09-10
郑州广毓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are many problems in the in vitro expansion of existing epidermal stem cells, such as the complex composition of the medium, the need for trophoblast co-cultivation, poor stability, etc., there are safety hazards in clinical application, and the cell proliferation rate is slow, and a sufficient number cannot be obtained in a short time Epidermal stem cells, etc., cells differentiate quickly, and generally can only be passaged to P3-P5 generations, which also limits the number of cells that can be expanded.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A medium for epidermal stem cell expansion, consisting of basal medium RPMI1640 and the following components added to RPMI1640 medium: thymosin α1, glycerol phosphatidylcholine, raffinose, ferrous glycinate, coenzyme Q10, vitamin E, and compound amino acids ;The final concentration of each component in the medium is: thymosin α1 2.8μg / mL, glycerol phosphatidylcholine 2.4μg / mL, raffinose 3.2ng / mL, ferrous glycinate 12.5ng / mL, coenzyme Q10 2.5ng / mL mL, vitamin E 15 μg / mL, glutamic acid 6 μg / mL.

[0028] The preparation method of the above-mentioned medium comprises the following steps: adding thymosin α1, glycerol phosphatidylcholine, raffinose, ferrous glycinate, coenzyme Q10, vitamin E, and compound amino acids to the basal medium, stirring evenly, and using a 0.22 μm filter to Sterilize by membrane filtration.

Embodiment 2

[0030] A medium for epidermal stem cell expansion, consisting of basal medium RPMI1640 and the following components added to RPMI1640 medium: thymosin α1, glycerol phosphatidylcholine, raffinose, ferrous glycinate, coenzyme Q10, vitamin E, and compound amino acids ;The final concentration of each component in the culture medium is: thymosin α1 2.5μg / mL, glycerol phosphatidylcholine 1.8μg / mL, raffinose 3.0ng / mL, ferrous glycinate 10ng / mL, coenzyme Q10 1.5ng / mL , vitamin E12μg / mL, arginine 2μg / mL, tryptophan 3μg / mL.

[0031] The preparation method of the above medium is the same as in Example 1.

Embodiment 3

[0033] A medium for epidermal stem cell expansion, consisting of basal medium RPMI1640 and the following components added to RPMI1640 medium: thymosin α1, glycerol phosphatidylcholine, raffinose, ferrous glycinate, coenzyme Q10, vitamin E, and compound amino acids ; The final concentration of each component in the medium is: thymosin α1 3.2 μg / mL, glycerol phosphatidylcholine 2.7 μg / mL, raffinose 5.5 ng / mL, ferrous glycinate 15 ng / mL, coenzyme Q10 3 ng / mL, Vitamin E 18μg / mL, lysine 8μg / mL.

[0034] The preparation method of the above medium is the same as in Example 1.

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Abstract

The invention discloses an epidermal stem cell amplification culture medium. The epidermal stem cell amplification culture medium comprises a basic culture medium and the following components added in the basic culture medium: thymosin alpha 1, glycerol phosphatidylcholine, raffinose, ferrous glycinate, coenzyme Q10, vitamin E and compound amino acid. According to the culture medium, trophoblasts do not need to be added for co-culture, the proliferation rate of epidermal stem cells can be remarkably increased by adding thymosin alpha 1 and glycerol phosphatidylcholine, and the number of cells obtained through culture is increased. And raffinose and ferrous glycinate are also added into the culture medium, so that the epidermal stem cells can maintain an undifferentiated state, the proliferation activity of the cells is maintained, and the subculture efficiency of the cells is improved. The invention further provides a preparation method of the epidermal stem cell amplification culture medium, and the preparation method is simple and convenient in process and easy to operate. The invention further provides application of the culture medium in epidermal stem cell amplification, in-vitro mass amplification of epidermal stem cells is achieved, and a large number of epidermal stem cells can be harvested within a short time.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to an epidermal stem cell expansion medium, a preparation method and application thereof. Background technique [0002] The skin is the largest organ that constitutes the human body. It is in direct contact with the external environment, and it protects the human body from various harmful factors including harmful microorganisms and external stimuli such as ultraviolet rays. An organ essential for the maintenance of required biochemical functions. There are many kinds of stem cells in the skin epidermis, among which epidermal stem cells exist in the outermost basal layer of the skin. It plays an important role in the formation of epidermis, the stability of epidermal tissue structure, the healing of skin wounds, and the occurrence and development of various skin diseases. [0003] With the development of biology, it is found that epidermal stem cells play an extremely important role in t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074
CPCC12N5/0625C12N2501/30C12N2500/36C12N2500/34C12N2500/32C12N2500/38C12N2501/999
Inventor 廖洪花贾定福
Owner 郑州广毓生物科技有限公司
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