Method for producing tissue engineering skin outside of the body with scarifiskin stem cell

A tissue-engineered skin and epidermal stem cell technology, applied in prosthesis, medical science and other directions, can solve the problems of lack and the influence of seed cell development, and achieve the effect of strong proliferation ability and high purity

Inactive Publication Date: 2008-04-02
陆洪光
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0007] 3. Type I collagen matrix is ​​used as a dermal substitute scaffold to cultivate artificial skin, which lacks the structure of artificial dermis and other necessary components of basement membrane (such as Vimentin, type IV collagen, laminin, etc.), which will have a certain impact on the development of seed cells. influences

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0035] Sources of reagents used in the examples of the present invention: DMEM medium, keratinocyte serum-free medium (KC-SFM), Ham's F12 nutrient solution, bovine pituitary extract, recombinant human epidermal growth factor (hEGF), non-essential amino acids, insulin All were purchased from Gibco Company of the United States; human placenta type IV collagen, Dispase enzyme, trypsin, cholera toxin, penicillin and streptomycin were all purchased from Sigma Company of the United States; recombinant human basic fibroblast growth factor (bFGF) was purchased from Zhuhai Yi Sheng company, fetal bovine serum (FBS) was purchased from Hangzhou Sijiqing Company, mouse anti-human keratin 19 (K19) monoclonal antibody, mouse anti-human β1-integrin (Integrin-β1) monoclonal antibody were purchased from Antibody Diagnostica Company of the United States, mouse Anti-human keratin monoclonal antibody was purchased from Dako Company of Denmark, and immunohistochemistry kit was purchased from Jingme...

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Abstract

The invention provides a method of manufacturing tissue engineering skin out of the body with epithelial stem cells. The method includes the following steps: (1) preparation of the epithelial stem cells: epithelial cells are made into cell suspension liquid, adhered and screened by IV collagen, and cultivated with epithelial stem cell media; (2) preparation of deepitheliarizing derma; (3) construction of the tissue engineering skin: epithelial stem cells for 2nd to 5th generation cultivation are selected, Dispase enzyme is adopted to digest the bossy cell patches in the size of rice to soya beam, the patches are inoculated to the deepitheliarizing derma, the tissue media is added, to cultivate the tissue engineering skin underwater or on the water. The tissue engineering skin constructed by the method of the invention is more suitable for the biological characteristics of human skin.

Description

technical field [0001] The invention relates to a method for preparing tissue-engineered skin, in particular to a method for making tissue-engineered skin in vitro with epidermal stem cells. Background technique [0002] In recent years, with the development of in vitro seed cell culture technology and dermis substitutes, people have developed artificial skins with various structures similar to human skin, which are called tissue engineered skins. Tissue engineered skin is not only used for transplantation and repair of skin after trauma (such as burns), but also for research on skin development, pathogenesis of skin diseases, and treatment of certain skin diseases. Therefore, it has broad development and application prospects in the medical field. Skin seed cells and dermal substitutes are important elements for constructing tissue-engineered skin. Since epidermal stem cells are pluripotent stem cells with the potential to proliferate and differentiate into epidermis and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/36
Inventor 陆洪光王新宇
Owner 陆洪光
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