小鼠精原干细胞分选

Sorting of Spermatogonial Stem Cells in Mice

目的：寻求富集小鼠精原干细胞的有效方法。方法：取6周龄雄性昆明白小鼠20只，手术制作成双侧隐睾模型，继续饲养2～3个月后，切取睾丸，用酶两步消化法制成单细胞悬液，加入FITC标记的抗α6-integrin抗体和PE标记的抗c—kit抗体冰浴20min后，利用流式细胞仅筛选出具有sidescatter^1o、α6-integrin^＋、c-kit^-特征的细胞，苔盼兰染色监测细胞活性。另取20只小鼠作为对照组，相同条件下饲养相同时间。结果：隐睾小鼠筛选出的精原干细胞占睾丸细胞总数的2．8％，95％以上的细胞具有活性。结论：利用免疫荧光激活细胞分选术能分离出大量有活性的精原干细胞。

Objective： To search for an effective method for enriching spermatogonial stem cells in mice. Methods ： Bilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-α6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and posi- tively stained for α6-integrin and negative c-kit expression. And the viability of the isolated ceils was assessed by trypan blue exclusion. Results ： The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable. Conclusion： FACS can be used to isolate quantities of viable spermatogonial stem ceils.