Regulation of the fetal human adrenal cortex: effects of adrenocorticotropin on growth and function of monolayer cultures of fetal and definitive zone cells

Abstract

Monolayer cultures have been prepared from both definitive and fetal zones of the human fetal adrenal cortex. Cultures from each zone consist predominately of adrenocortical cells, as determined by a specific morphological retraction response to ACTH, and by ACTH-induced inhibition of DNA synthesis and cell proliferation. Cell growth was stimulated by fibroblast growth factor. ACTH stimulated steroidogenesis in cells from each zone with an ED50 of 0.4--1.0 nM and at a maximal concentration of 5 nM. Short term stimulation of less than 24 h with ACTH produced a pattern of steroid secretion that was characteristic of the zone of origin. Definitive zone cultures produced both cortisol and dehydroepiandrosterone plus its sulfate (DHA/S), with cortisol production exceeding DHA/S production. Fetal zone cultures produced more DHA/S than cortisol. 3 beta-Hydroxysteroid dehydrogenase, delta 4,5-isomerase enzyme activity was 3-fold less in fetal than in definitive zone cultures. Long term stimulation of 1--4 days with ACTH, 8-bromo-cAMP, or cholera toxin increased steroidogenic capacity in cultures from both zones. The pattern of steroid production by definitive zone cells remained constant, but cortisol production was preferentially increased in fetal zone cells. Forty-eight-hour treatment of fetal zone cells with ACTH increased 3 beta-hydroxysteroid dehydrogenase, delta 4,5-isomerase activity 5-fold. alpha-, beta-, gamma 1-, gamma 2-, and gamma 3-MSH were not effective steroidogenic agents for either zone. These studies indicate that steroidogenic agents induce in fetal zone cells steroid production characteristic of definitive and adult adrenocortical cells.