Primary culture, identification and functional study of rat Leydig cells.
- Author:
Feng YING
1
;
Yi GONG
;
Jia-Yin SUN
;
Jie SHEN
;
Xiao-Dong HAN
Author Information
- Publication Type:Journal Article
- MeSH: 3-Hydroxysteroid Dehydrogenases; metabolism; Animals; Cell Culture Techniques; Cell Separation; methods; Cells, Cultured; Centrifugation, Density Gradient; methods; Histocytochemistry; Leydig Cells; cytology; enzymology; physiology; Male; Rats; Rats, Sprague-Dawley
- From: National Journal of Andrology 2008;14(1):7-10
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo set up a stable primary culture system of Leydig cells with higher purity.
METHODSWe 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.
RESULTSThe 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.
CONCLUSIONLeydig 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.
