A model for in vitro studies of epidermal homeostasis: proliferation and involucrin synthesis by cultured human keratinocytes during recovery after stripping off the suprabasal layers

Abstract

In normal epidermis there is a balance between the rate of cell division and the rate of terminal differentiation. This can be perturbed by injuries such as wounding or tape-stripping, but is reestablished during recovery. The aim of the studies described in this report was to assess whether cultures of human keratinocytes could be used as an experimental model for investigating the mechanism by which epidermal homeostasis is established and maintained. The suprabasal layers were stripped from confluent keratinocyte cultures by incubation in low-calcium medium (0.1 mM calcium ions). After return to normal medium (2 mM calcium ions), the basal layer regenerated a stratified culture of approximately the same thickness as controls. The kinetics of proliferation and terminal differentiation were monitored by measuring the total number of cells and proportion of involucrin-positive cells at intervals before, during, and after stripping. During recovery the proportion of cells expressing involucrin, assessed by immunofluorescence microscopy and polyacrylamide gel electrophoresis, rapidly returned to control levels, but the total number of cells per dish rose more slowly and often failed to reach control values. Thus, terminal differentiation was initially stimulated at the expense of proliferation. Our in vitro model of epidermal regeneration should provide a useful complement to intact skin and animal models for analyzing epidermal homeostasis.