[Primary culture, identification and functional study of rat Leydig cells].
OBJECTIVE
To set up a stable primary culture system of Leydig cells with higher purity.


METHODS
We separated Leydig cells from other testicular cells, such as Sertoli and germ cells, by enzymatic digestion in combination with Percoll density gradient centrifugation and identified Leydig cells by 3beta-HSD staining.


RESULTS
The purity achieved by this method was above 95% and the total number of Leydig cells obtained from one testicle was about 1 x 10(6). The cytoplasm of Leydig cells was stained in deep blue by 3beta-HSD staining, and these cells possessed testosterone-secreting capability.


CONCLUSIONS
Leydig cells can be separated by enzymatic digestion combined with Percoll density gradient centrifugation, and 3beta-HSD staining to identify Leydig cells is simple and feasible with high purity.
Ying
Feng
F
Laboratory of Immunology and Reproductive Biology, Nanjing University School of Medicine, Nanjing, Jiangsu 210093, China.
Gong
Yi
Y
Sun
Jia-Yin
JY
Shen
Jie
J
Han
Xiao-Dong
XD
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods; D006651:Histocytochemistry; D007985:Leydig Cells / Q000166:cytology* / Q000201:enzymology / Q000502:physiology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D015096:3-Hydroxysteroid Dehydrogenases