Differentiation and barrier formation of a cultured composite skin graft

The differentiation and barrier formation of cultured composite skin grafts (CSGs) were assessed by histology and measurements of surface electrical capacitance (SEC) in vitro and in vivo. Keratinocytes cultured on the surface of acellular dermis were lifted to the air-liquid interface and analyzed for 30 days in vitro. Initially, SEC measurements of CSGs (n = 11) were high but quickly dropped between days 4 and 6 and remained steady for 30 days, indicating barrier formation by the epidermis. Histology of the CSGs (n = 6) demonstrated stratification of the epidermal cells and partial formation of the stratum corneum by day 3 that was complete by day 7. CSGs (n = 5) were transplanted to athymic mice, where they formed a stratified and differentiated epidermis. SEC measurements of CSGs remained low after transplant, suggesting that exposure to the air-liquid interface improved the maturation of CSGs in vitro prior to transplant.