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Updated: Jul 7, 2026
Culturing Primary Rat Inner Medullary Collecting Duct Cells
Published on: June 21, 2013
[Primary culture, identification and functional study of rat Leydig cells]
Feng Ying1, Yi Gong, Jia-Yin Sun
1Laboratory of Immunology and Reproductive Biology, Nanjing University School of Medicine, Nanjing, Jiangsu 210093, China.
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.
Conclusion:
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.
