LATERAL GROWTH AND TERMINAL DIFFERENTIATION DURING REPEATED EPIDERMAL REGENERATION INVITRO - AGE DEPENDENCE AND MODULATION BY CHOLERA-TOXIN

By incubating multilayered primary cultures of human epidermal keratinocytes in a low calcium medium, the suprabasal layers can be stripped off leaving a basal cell monolayer. When this monolayer is refed normal calcium medium a reproducible series of cell kinetic, morphological and biochemical changes take place resulting in the regeneration of a multilayered tissue. The stripping procedure seems to induce the selective proliferation of a cohort of basal cells that is committed to vertical migration and rapid terminal differentiation. In contrast, when the basal cells are allowed to regenerate in the presence of the strong mitogen, cholera toxin, lateral growth and continued proliferation are favoured at the expense of the capacity of the cells to differentiate. Repeated stripping of the same cultures disclosed a considerable heterogeneity in the capacity of the basal cells to regenerate the suprabasal layers. The number of times the basal cells could restore the suprabasal layers after repeated stripping varied from four to nine times. A negative correlation between donor age and regenerative capacity was observed. The experiments with repeated stripping of the same cultures also showed that the capacity to proliferate and to restore the multilayering was fully retained for at least four cycles of stripping-regeneration, whereas the capacity to terminally differentiate was rapidly lost. It is suggested that the present system of regenerating epidermal tissue cultures may serve as an experimental model for the study of epidermal tissue homeostasis and cellular aging.