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Culture fluid for promoting formation of tissue engineering skin lipid and formulating method thereof

A tissue-engineered skin and culture fluid technology is applied to the culture fluid and its preparation field for promoting the formation of tissue-engineered skin lipid structures, and can solve the problems of high misjudgment rate, cumbersome operation during use, inconsistent data, etc. The ability of structure formation, the simple preparation method and the effect of strong stability

Inactive Publication Date: 2009-07-22
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the stratum corneum is not fully matured and differentiated, and has not formed a good lipid structure, the toxicity of the detected compound to active cells is inconsistent with the data of its own toxicity in vivo, resulting in a high misjudgment rate and the required prediction accuracy cannot be achieved. Spend
[0004] At present, there is no commercial culture medium on the market that promotes the formation of lipid structure in tissue engineered skin. The culture medium used in the research of tissue engineered skin in the existing laboratory has the advantages of reagent stability, cell culture effect and promotion of keratinocyte differentiation. There are deficiencies in capacity, and the use process is cumbersome
After searching, it has been found that there is a culture medium with high stability, easy operation, cell culture effect and excellent ability to promote the formation of tissue-engineered skin lipid structure, but no public information has been found so far.

Method used

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Embodiment Construction

[0043] For further illustrating the present invention, specifically set forth in conjunction with the following examples:

[0044] 1. Preparation of culture medium:

[0045] Its components and the consumption of each component are:

[0046] (1) Basal culture medium: mixed with the following substances in volume percentage:

[0047] DMEM culture medium 65%

[0048] F12 medium 25%

[0049] Fetal Bovine Serum 10%

[0050] (2) Nutrients:

[0051] Hydrocortisone 0.4~0.5μg / mL

[0052] Epidermal growth factor 5~15ng / mL

[0053] Isoproterenol 1~3×10 -6 mol / L

[0054]Insulin 5~15μg / mL

[0055] Transferrin 5~10μg / mL

[0056] Triiodothyronine 2~6×10 -9 mol / L

[0057] Adenine 1~3×10 -4 mol / L

[0058] Vitamin C 50~100μg / mL

[0059] Retinoic acid 4~10μg / mL

[0060] Cholera toxin 0.1~1nmol / L

[0061] L-serine 1~3×10 -2 mol / L

[0062] L-carnitine 1~5×10 -5 mol / L

[0063] Palmitic acid 15~35μmol / L

[0064] Linoleic acid 5~25μmol / L

[0065] Arachidonic acid 5~10μmol / L

...

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Abstract

The invention discloses a culture fluid that promotes the generation of a lipid structure of tissue engineered skin. A basic culture fluid is added with hydrocortisone, epidermal growth factors, isoproterenol, insulin, transferrin, triiodothyronine, adenine, vitamin C, tretinoin, cholera toxin, L-serine, L-novain, palmic acid, linoleic acid, arachidonic acid, bovine pituitary extract, bovine serum albumin, calcium chloride, penicillin, streptomycin and fungizone B, etc. Tissue engineered skin with mature and completely differentiated horny layer and good lipid structure can be obtained by cultivating keratinocyte in the culture fluid of the invention, the culture fluid has strong stability, long shelf life of about 12 months, simple and convenient preparation method, and convenient product synthesis and mass production.

Description

technical field [0001] The invention relates to biological reagents in the research of tissue-engineered skin, in particular to a culture solution for promoting the formation of tissue-engineered skin lipid structure and a preparation method thereof. Background technique [0002] The skin is the largest organ of the human body, and it consists of three parts, namely the epidermis, dermis and subcutaneous layer. The epidermis is located in the outermost layer of the skin, with a thickness between 0.06 and 0.8 mm. Its microstructure is composed of basal layer, spinous cell layer, granular cell layer and stratum corneum. The stratum corneum is the final product of keratinocyte differentiation, 75% to 80% of its composition is protein, 5% to 15% is lipid, and a structure with a thickness of about 10 μm is composed of 10 to 15 layers of cells. Many important physiological functions of the stratum corneum are related to the lipid structure, which can effectively prevent the damag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61L27/60C12N5/071
Inventor 邹运动董益阳程艳王军兵
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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