Çulturing of kerationocytes in serum-free medium

Background and To establish a simple method for preparation of pure epidermal cell in serum-free medium, without co culturing with lethally irradiated 3T3 cells. Â piece of skin biopsy was taken from a rat. Âfter trypsinazation to separate epidermal layer, the epidermis was cut into explants 1×1 mm and were laid at 50 mL on tissue cultured flasks. Then, the explants were covered with serum-free keratinocyte growth medium, while the culture medium was changed every two days. The keratinocyte colonies were expanded in flasks and proliferated to 70% confluency on day 21. Âfter two subcultures, the cells were frozen. Pure epidermal cells are established by this technique in a serum-free keratinocyte growth medium, under feeder cell-free condition.Çonclusion: Çulturing of keratinocytes in serum-free medium is proved to be a useful and simple method for keratinocyte isolation. The successful culturing of keratinocytes from a small skin biopsy can be useful in the treatment of major burn wounds.