Terminal Epidermal Differentiation of Human Keratinocytes Grown in Chemically Defined Medium on Inert Filter Substrates at the Air-Liquid Interface

摘要: A fully differentiated epithelium having the features of epidermis was obtained in vitro by culturing second-passage normal human keratinocytes (NHK) chemically defined medium MCDB 153 on inert filter substrates at air-liquid interface for 14 d. Vertical sections stained histology and indirect immunofluorescence studies show a correct stratification expression differentiation markers. The presence desmosomes, keratohyalin granules, lamellar formation more than ten-layers stratum corneum evidenced electron microscopy. Moreover, lipids typical were present these cultures, including ceramides, which are thought to be responsible relative impermeability corneum. Under our culture conditions, i.e., interface, use de-epidermized dermis as substrate did not stimulate keratinocyte acetate cellulose or polycarbonate membrane substrates. obtaining well-differentiated grown filters should useful standard system studying epidermal differentiation, re-epidermization, cytotoxicity, permeation, transepidermal drug delivery.