title: Preliminary observations of bovine adrenal fasciculata-reticularis cells in monolayer culture: steroidogenesis, effect of ACTH and cyclic AMP.
abstract: Bovine adrenocortical cells dispersed by trypsin digestion of fasciculata-reticularis minces were maintained in monolayer culture for up to 6 weeks. During the first week cells grown in medium containing ACTH (1 mU/ml) secreted steroids at a rate 10 to 20-fold greater than control cultures, cortisol accounting for 80-90% of the corticotrophic response. Using tracer amounts of [3H] progesterone and [3H] pregneolone, the major products were cortisol, corticosterone, 11-deoxycortisol and 11-deoxycorticosterone in decreasing order of magnitude. After 10 to 15 days in culture steroidogenesis was no longer enhanced by ACTH. This was concomitant with an apparent loss of 11 beta-hydroxylase activity which was mainly manifested by a sharp increase in the formation of 44-deoxycortisol. Short-term incubations of these cells during the first week in culture provided evidence that steroidogenesis was related to ACTH concentrations (from 0.1 to 100 muU/ml) and stimulated by dibutyryl cyclic AMP, the corticotrophic responses being further enhanced by theophylline (0.5to 50 mumoles/5 ml). Exposure of the cells to ACTH (50 muU/ml) resulted in a rapid increase in intracellular cyclic AMP contractions concomitant with a progressive increase in the corticosteroids released into the medium.
mesh_terms: D000302:Adrenal Cortex / Q000166:cytology / Q000378:metabolism*; D000311:Adrenal Glands / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001681:Biological Assay; D003994:Bucladesine / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D003345:Corticosterone / Q000096:biosynthesis; D003350:Cortodoxone / Q000096:biosynthesis; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D005938:Glucocorticoids / Q000096:biosynthesis*; D006854:Hydrocortisone / Q000096:biosynthesis; D011284:Pregnenolone / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D013997:Time Factors
chemical_list: D003470:Culture Media; D005938:Glucocorticoids; D011374:Progesterone; D003994:Bucladesine; D011284:Pregnenolone; D000324:Adrenocorticotropic Hormone; D000242:Cyclic AMP; D003345:Corticosterone; D003350:Cortodoxone; D006854:Hydrocortisone