Abstract: The present invention relates to the treatment of skin defect by organotypically cultured autologous keratinocytes isolated from the outer root sheath of hair follicles. Methods for primary as well as subsequent organotypic cultures (epidermal equivalents) in fully defined media eventually supplemented by autologous human serum and substances isolated from blood components, with minimal allogeneic biological supplements, are disclosed. Techniques to prepare epidermal equivalents for transplantation are included, as well as a method for the transport of the epidermal equivalents.