摘要
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.
出处
《中华男科学杂志》
CAS
CSCD
2007年第1期24-26,共3页
National Journal of Andrology
基金
湖北省教育厅资助项目(2000B03008)
武汉大学青年博士创新基金项目
关键词
精原干细胞
免疫荧光激活细胞分选术
小鼠
spermatogonial stem cells
fluorescence-activated cell sorting
mouse