Cultivation,frozen storage,and clonal growth of normal human epidermal keratinocytes in serum-free media

Summary Methods are described for serum-free culture of human epidermal keratinocytes derived from neonatal foreskin tissue. Cultures are initiated, stored frozen, and returned to active growth, all with bovine pituitary extract as the only undefined supplement. Clonal growth assays are then performed in a biochemically defined medium. The degree of stratification and differentiation in the defined medium (and also with pituitary extract) is controlled by the extracellular calcium ion concentration.