Separation and culturing method of human epidermis stem cell
A technology of epidermal stem cells and epidermal cells, which is applied in the field of isolation and in vitro culture of human epidermal stem cells, and can solve the problems that the characteristics of epidermal stem cells cannot be guaranteed
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[0017] The foreskin skin of (human) healthy male after circumcision was taken to prepare epidermal cells and fibroblasts.
[0018] 1. Separation of epidermal cells (keratinocytes): take a skin 1-2 cm long and about 2 mm wide, wash it 3 times with PBS containing antibiotics; put it in protease solution, digest it at 4°C for 16 hours; take out the skin, peel off the epidermis and Dermis layer: collect epidermis slices, place in 0.25% trypsin / 0.02% EDTA (1:1) mixture, digest at 37°C for 10-15 minutes, stop digestion, pipette slightly and filter, collect cell suspension, centrifuge and discard Supernatant, add fresh culture medium, resuspend cells, adjust cell concentration to 1×10 3 / ml.
[0019] 2. Preparation of trophoblast cells: collect the above-mentioned dermis slices after peeling off the epidermis and dermis, place them in 625U / ml collagenase solution, collect the fibroblast suspension after digestion, and culture the fibroblasts until they spread about 80%. Add mitomyc...
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