Steroidogenesis of cultured purified pig Leydig cells: secretion and effects of estrogens.

Using a primary culture of purified immature pig Leydig cells we have demonstrated: (1) that during the first 3 days of culture there is a 'spontaneous' maturation of the steroidogenic response to hCG, as expressed by a 50-fold increase of the steroidogenic capacity, an increased secretion of both dehydroepiandrosterone sulfate (DHAS) and testosterone (T), a shift of the DHAS/T ratio (5 on day 0 vs. 0.5 on day 3) without significant changes in the number of hCG-binding sites; (2) that purified cells, on day 3 following hCG stimulation, secrete large amounts of T and small amounts of E2 (T/E2 congruent to 150) and detectable amounts of estrone, while crude pig interstitial cells under the same conditions secrete less T but 40-50 times more estrogens (T/estrogens congruent to 1.5); (3) that the steroidogenic responsiveness of purified Leydig cells is not impaired by E2 treatment, in spite of the fact that Leydig cells contain specific estradiol receptors (approximately 10000 sites/cell). These data suggest that in this model the main source of testicular estrogens are not Leydig cell but some other testicular cell types, and that the lack of effect of estrogens on pig Leydig cell steroidogenesis is not due to absence of estrogen receptors.