Enzymatic liberation of viable cells of human skin
Hentzer, B.; Kobayasi, T.
Acta Dermato-Venereologica 58(3): 197-202
1978
ISSN/ISBN: 0001-5555 PMID: 78632 Document Number: 132002
To obtain viable cells from normal human skin, clostridial collagenase was used. Crude collagenase digestion of collagen fibers and basal lamina results in free dermal cells and sheets of epidermis. The collagenase was tested at various concentrations, solvents and incubation periods. The specimens digested were either split or full thickness skin of varying size. The optimal result was obtained by using small (3 mm across) split skin pieces incubated in 2 mg/ml collagenase. The choice of solvent MEM , MEM supplemented with serum and Tris buffer, was less important. After 3 h incubation the epidermis was peeled off in sheets and finally dissociated by trypsin-EDTA. The corium was completely digested after 6 h. After 6 h incubation no viable cells were seen. The epidermal cells appeared mainly as polygonal cells of various sizes and a few little dendritic cells. The dermal cells had a heterogeneous morphology during the 1st wk of cultivation. After 2 wk the cells appeared as fibroblast-like cells.