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
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[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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