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The transfer of pigment granules from melanocytes to keratinocytes was studied using a new living skin equivalent (SE) model in vitro. The model was constructed by plating human neonatal melanocytes onto a dermal equivalent (DE) before it was overgrown with keratinocytes. The dermal component of the SE arises in vitro through the action of fibroblasts, which compact matrix proteins into a tissue. It becomes keratinized as keratinocytes migrate out of 2-mm punch biopsies of human neonatal skin embedded in the DE; keratinocytes from the biopsies covered the lattice in 14 days. A basal lamina develops at the dermal-epidermal junction in vitro. Exposure of some SEs to UVB irradiation for 14 days stimulated and enhanced pigment transfer. Pigment transfer from melanocytes to keratinocytes was documented in light and electron microscopic studies. Melanosomes, identified by their pigment as well as by dopa oxidase staining, were dispersed throughout the keratinocyte cytoplasm. We conclude that the SE model is valuable for studying the relationship between melanocytes and keratinocytes in vitro; since the SE has been shown to serve as a skin replacement, pigmenting it may be expected to increase its usefulness.
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