title: The functional activity of adult mouse Leydig cells in monolayer culture. Effect of lutropin and foetal calf serum.
abstract: Purified Leydig cells were obtained from adult mouse testes by mechanical dispersion followed by Percoll density-gradient centrifugation as described by Schumacher et al. (1978). The cells were then established in monolayer culture by maintaining them in medium and 10% serum at 32 degrees C in 95% O2, 5% CO2. The cells rapidly attached to the culture dishes, gradually flattened and became epitheloid in appearance. Testosterone production by the cells in response to maximum stimulating levels of LH (100 ng/ml) and dibutyryl cyclic AMP (1 mM) was maintained for at least 2 days (approximately 1 microgram/10(6) cells/2h) and then declined to lower levels by days 3-4. Cyclic AMP production in response to LH was higher on day 1 than day 0 and then declined to lower levels by days 3-4. Binding of [125I]hCG was similar on day 0 and day 1 (approximately 20 fmoles/10(6) cells) and then declined to lower levels by days 3-4. The functional activity of the cells cultured in 0, 1 and 10% foetal calf serum was also examined; no significant effect of the serum on LH-stimulated testosterone or cyclic AMP production was found; however, a decrease of up to 50% in the binding of [125I]hCG to the Leydig cells occurred in the presence of serum. These results demonstrate that the function of differentiated adult Leydig cells can be maintained for at least 2 days in culture.
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D001769:Blood; D002417:Cattle; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000378:metabolism; D003470:Culture Media; D000242:Cyclic AMP / Q000096:biosynthesis / Q000494:pharmacology; D007985:Leydig Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007986:Luteinizing Hormone / Q000494:pharmacology*; D008297:Male; D051379:Mice; D013739:Testosterone / Q000096:biosynthesis
chemical_list: D006063:Chorionic Gonadotropin; D003470:Culture Media; D013739:Testosterone; D007986:Luteinizing Hormone; D000242:Cyclic AMP; D015056:1-Methyl-3-isobutylxanthine