成年小鼠睾丸间质细胞的分离、鉴定及功能检测

Isolation,Identification and Function Test of Adult Mouse Leydig Cells

为建立成年小鼠睾丸间质细胞分离纯化方法,并对分离纯化的睾丸间质细胞进行睾酮分泌功能检测,对成年小鼠睾丸组织进行I型胶原酶消化、Percoll分离液密度梯度离心,分离成年小鼠睾丸间质细胞。细胞纯度用3β-羟类固醇脱氢酶(3β-hydroxy-steroid-dehydrogenase,3β-HSD)染色鉴定。将分离纯化的睾丸间质细胞进行体外培养,在基础睾酮和人绒毛膜促性腺激素(hCG)刺激培养条件下,用放射免疫分析法对培养上清中的睾酮含量进行检测。结果显示,通过该方法能获得高纯度成年小鼠睾丸间质细胞(>95%),培养的睾丸间质细胞具有分泌基础睾酮以及对hCG刺激反应的能力。提示采用该方法对成年小鼠睾丸间质细胞进行分离纯化具有高效性、可用性,通过该方法获得的睾丸间质细胞是体外研究药物对睾酮分泌功能影响的良好模型。

In order to establish the method for ability of Leydig cell testosterone secretion, isolating and purifying adult mouse Leydig cells, and test the adult mouse testes were digested with type I collagenase and were then purified by Percoll gradient centrifugation. The cell purity was determined using 3β-hydroxyste- roid dehydrogenase （3β-HSD） staining method. Isolated and purified adult mouse Leydig cells were cul- tured under basal and hCG stimulated conditions, and testosterone concentrations in the media were then determined using RIA method. The results reveled that highly purified （〉95G） 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.