In vitro fabrication of human epidermal tissues that mimic the biochemical and morphologic properties of human skin, known as skin-equivalent (organotypic) cultures, has opened new avenues in the study of skin biology. In this chapter, methods for the generation of these tissues from their component parts are described. Conditions for culture of human keratinocytes and fibroblasts that allow optimal growth in skin equivalent cultures are delineated. These cell types are then sequentially combined so that keratinocytes are grown at an air-liquid interface on a contracted collagen gel containing dermal fibroblasts. The methods described enable the generation of human epidermal tissues that show in vivo-like tissue architecture and phenotype.
