Preparation method of tissue engineering skin containing appendant organs

A technique of tissue engineering skin and tissue engineering, which is applied in the field of preparation of tissue engineering skin containing appendages, can solve the problems of not being able to function normally in the epidermis, troubled tissue engineering skin, and increased fragility of the epidermis, and achieves vigorous cell metabolism , Facilitate large-scale production, low immune rejection effect

Inactive Publication Date: 2010-07-14
FOURTH MILITARY MEDICAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Because epidermal cells tend to lose their keratinization characteristics during in vitro culture, the epidermal layer of the prepared tissue-engine...

Method used

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Examples

Experimental program
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Effect test

example 1

[0029] Step 1. Preparation of culture medium and induction of cell orientation

[0030] Preparation of culture medium A (induction medium for amnion epithelial cells to keratinocytes), its components include: commercial medium DMEM 360ml, commercial medium F12 90ml, fetal bovine serum 50ml, adenine 200mM, insulin 2μg / ml, transferrin 6μg / ml, epidermal growth factor 15ng / ml, hydrocortisone 0.2μg / ml;

[0031] Prepare culture medium B (medium for mesenchymal stem cells to induce dermal papilla), its components include: 300ml of commercial medium DMEM, 125ml of commercial medium F12, 75ml of fetal bovine serum, 150ng / ml of wnt3a protein, Fibroblast growth factor 2 (FGF-2) 4ng / ml;

[0032] Prepare culture medium C (induction medium from amniotic membrane epithelial cells to sweat gland epithelial cells), its components include: 450 ml of commercial medium DMEM, 50 ml of fetal bovine serum, 12 ng / ml of nerve growth factor (NGF), epidermal growth factor Factor 15ng / ml, transforming...

Embodiment 2

[0044] Step 1. Preparation of culture medium and induction of cell orientation

[0045] Prepare culture medium A (medium for inducing amnion epithelial cells to keratinocytes), its components include: commercial medium DMEM 360ml, commercial medium F12 90ml, fetal bovine serum 50ml, adenine 260mM, insulin 5μg / ml, transferrin 5μg / ml, epidermal growth factor 13ng / ml, hydrocortisone 0.4μg / ml;

[0046] Prepare culture medium B (medium for mesenchymal stem cells to induce dermal papilla), its components include: 300ml of commercial medium DMEM, 125ml of commercial medium F12, 75ml of fetal calf serum, 130ng / ml of wnt3a protein, Fibroblast growth factor 2 (FGF-2) 8ng / ml;

[0047] Prepare culture medium C (induction medium from amniotic membrane epithelial cells to sweat gland epithelial-like cells), its components include: commercial medium DMEM 450ml, fetal bovine serum 50ml, nerve growth factor (NGF) 15ng / ml, epidermal growth factor Factor 10ng / ml, transforming growth factor (T...

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Abstract

The invention relates to a preparation method of tissue engineering skin containing appendant organs. In the invention, the preparation method comprises the following steps of: inoculating amniotic mesenchyme stem cells, amniotic mesenchyme stem cells induced to the directions of hair papillae, amniotic epithelial cells and amniotic epithelial cells induced to the directions of the epithelia of the sweat gland into a gel solution, then inoculating the amniotic epithelial cells and the amniotic epithelial cells induced by keratinocyte in a warp direction on the surface of a structure of the hair follicle and the sweat gland after culturing by a dermal layer by induced culture forming the structure of the hair follicle and the sweat gland, and obtaining the tissue engineering skin with the structure of the hair follicle and the sweat gland after culturing. The skin has elasticity, toughness, vigorous cellular metabolism, strong multiplication capacity, sufficient secretion of extracellular matrices, tight linkage of the cuticular layer, the inophragma and the cells of the dermal layer and little possibility of falling off, enhances the success rate of transplantation, can promote the healing of a wound surface, enhances the effects of restoration and replacement and has the function of regulating the immunological rejection of a receptor; by the contained structure of the hair follicle and the sweat gland, the function of the skin is more complete; the adopted amniotic tissues are postpartum wastes and have extensive source, strong cell multiplication capability and multiplepassage frequency; and the invention is beneficial to the mass production of seed cells and lowers the industrialization cost.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering of medical biological materials, and in particular relates to a method for constructing tissue-engineered skin with appendages by using cells derived from amniotic membrane. Background technique [0002] The skin is the largest organ of the human body. It has the functions of protecting the body, regulating body temperature, and excreting waste. It is an important tissue that covers and protects the body surface and internal organs. Defects and abnormalities of the skin and mucous membranes are often caused by inflammation, ulcers, trauma, burns, postoperative tumors, and congenital malformations. Normal skin contains a well-differentiated epidermis and dermis. An ideal healing wound should have a complete epidermis, dermis, hair follicles, sweat glands and other appendage structures; the epidermis can act as a barrier and protect, preventing harmful substances from passing through the ...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/38A61L27/36A61L27/60
CPCC12N2502/094C12N5/0698C12N2502/1323A61L27/38A61L27/60
Inventor 金岩杨鹭张勇杰王爱军
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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