New techniques for the in vitro culture of human skin keratinocytes and perspectives on their use for grafting of patients with extensive burns.

Human keratinocytes can be cultured in vitro and used for autografting of the injured epidermis. In this article, we describe our culture methods for propagating autologous keratinocytes for grafting, and we present an overview of the questions raised by the use of such methods. Our culture method involves a two-phase technique. Phase 1 is performed in serum-free medium and yields large quantities of a homogeneous, highly proliferative, basal cell-like population. Phase 2 involves the induced formation of cohesive, stratified sheets of differentiated keratinocytes resembling normal epidermis that can be used for grafting. This two-phase technique was used for preparation of autografts for full-thickness wounds of a massively burned patient. Important questions about the functional potential of cultured keratinocytes and the long-term behavior of cultured epidermal autografts in vivo remain to be answered.