The modified method for isolation and culture of highly homogeneous Leydig cell population from rat testes.
A modified method for isolation and culture of a pure population of rat Leydig cells is described. For obtaining crude interstitial cell suspension, decapsulated testes were dispersed in 0.02% collagenase solution in Ca2+, Mg2+--free Hanks medium for 1 hour. Then, approx. 5 X 10(7) cells were centrifuged in 10-90% discontinuous, isoosmotic Percoll gradient at 3000 g for 20 min. The cells from eight fractions obtained were collected and cultured in Eagle's MEM for 4 days. Using morphological methods, 1.059-1.070 g/ml density fraction contained 97% and 1.070-1.080 g/ml fraction contained 90% viable Leydig cells. The cells secreted testosterone to the culture medium and responded to LH stimulation with over four-fold increase in hormone secretion.
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000592:standards; D007985:Leydig Cells / Q000166:cytology* / Q000648:ultrastructure; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains