Production of basement membrane components by a reconstructed epidermis cultured in the absence of serum and dermal factors.

Abstract

A fully differentiated epithelium displaying features of human epidermis was obtained in vitro by culturing second-passage normal human keratinocytes for 14 days in defined medium and on an inert polycarbonate filter substratum at the air-liquid interface. Vertical sections stained for histology and indirect immunofluorescence studies showed that the 'basal' cells synthesize and secrete all major markers of hemidesmosomes and the lamina lucida. Components of the lamina densa are also expressed. Collagen VII is synthesized, but not secreted. Ultrastructural studies showed the presence of hemidesmosomes with major dense plaques and anchoring filaments, and a basement membrane-like structure was clearly identified. These results show that epidermal cells are able to produce hemidesmosomes and to secrete the major components of the dermo-epidermal junction in the absence of serum and dermal factors, suggesting that basement membrane synthesis and hemidesmosome assembly are not dependent on the presence of dermis.