Preparation method of tissue engineering skin containing appendant organs
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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: 2013-10-23
FOURTH MILITARY MEDICAL UNIVERSITY +1
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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-engineered skin becomes more brittle and cannot function normally. This is also a technical problem that has plagued the field of tissue-engineered skin for many years.
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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 present invention discloses an appendage-carrying tissue engineering skin producing method. The method involves inoculating amniotic mesenchymal stem cells, amniotic mesenchymal stem cells induced to the direction of dermal papilla, amniotic epithelial cells, amniotic epithelial cells induced to the direction of sweat gland epithelium in gel solution, inducing and culturing to form hair follicle and sweat gland structures after culturing of dermis layer, inoculating amniotic epithelial cells and amniotic epithelial cells induced to the direction of keratinocyte on the surface of said culture, culturing and obtaining tissue engineering skin carrying hair follicle and sweat gland structures. The skin is of elasticity, flexibility, strong cellular metabolism, strong proliferation ability and sufficient secretion of extracellular matrix. The epidermal layer is connected with the basal lamina and dermis layer cells tightly and is not easy to fall off. This increases the success rate of implantation; wound can be healed rapidly and repairing and substituting effects are improved. The tissue engineering skin has the function of regulating receptor immune rejection. The hair follicle and sweat gland structures carried make the skin to have more complete functions. The amniotic tissue is childbearing waste with wide range of sources. The cells of amniotic tissue have the capacity of strong proliferation and can be passaged for many times. The method helps mass production of seed cells and reduces industrial 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 ...
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