Isolation,Identification and Function Test of Adult Mouse Leydig Cells

In order to establish the method for isolating and purifying adult mouse Leydig cells,and test the ability of Leydig cell testosterone secretion,adult mouse testes were digested with type I collagenase and were then purified by Percoll gradient centrifugation.The cell purity was determined using 3β-hydroxysteroid dehydrogenase(3β-HSD)staining method.Isolated and purified adult mouse Leydig cells were cultured under basal and hCG stimulated conditions,and testosterone concentrations in the media were then determined using RIA method.The results reveled that highly purified(95%)adult mouse Leydig cells were obtained and cultured Leydig cells had the ability to produce basal testosterone and respond to human chorionic gonadotropin(hCG)stimulation.This suggests that the method for isolating and purifing adult mouse Leydig cells is highly efficient and usable,and Leydig cell obtained by this method is a good model to study in vitro drug function on testosterone secretion.