Primary culture, identification and functional study of rat Leydig cells / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To set up a stable primary culture system of Leydig cells with higher purity.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>