新生小鼠精原细胞分离和纯化的实验研究

Experimental study on the separation and purification of spermatogonia in neonatal mouse

目的:分离和纯化7～8d新生雄性小鼠精原细胞,为深入研究生精机理及其影响因素提供细胞来源和技术支持。方法:采用组合酶消化法制备7～8d雄性小鼠的生精细胞悬液;Percoll密度梯度离心法分离精原细胞,贴壁培养法进一步纯化精原细胞;滴片进行碱性磷酸酶染色;取同时间段雄性小鼠睾丸组织冰冻切片进行碱性磷酸酶染色。结果:精原细胞主要分布于45%～55%梯度间的Percoll中,经贴壁培养后纯度达75.2%。结论:用上述方法成功地分离和纯化了小鼠精原细胞。

Objective： To study the mechanism of spermatogenesis and its influence factors, the spermatogonia had to be separated and purified to provide technological groundwork and cells source. Method： Continuous enzymatic digestion was used to prepare sperrnatogenic cell suspension of male mouse of 7～8 days; Percoll density gradient centrifugation combined with plating culture method was used to isolate and purify spermatogonia ; alkaline phosphotase of the cells suspension drop and the testis tissues of the same time were detected. Results： Spermatogonia were mainly distributed in Percoll gradient between 45%～55%, purity of spermatogonia was 75.2% after being purified. Conclusion： Percoll density gradient centrifugation combined with continuous enzymatic digestion and plating culture method can isolate and purify spermatogonia from mouse 7～8 days postnatal.