Percoll非连续梯度法分离未成年绵羊精原细胞的研究  

Study of separating immature spermatogonia cells of sheep testis by discontinuous Percoll density

作　　者：武小虎[1] 张勇[1] 赵兴绪[1] 赵国顺[1] 马艳粉[1] 王朝凤[1] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《甘肃农业大学学报》2012年第6期24-29,共6页Journal of Gansu Agricultural University

基　　金：科技部国际合作重点项目计划(2005DFA30720)

摘　　要：为探索一种简单可行的分离未成年绵羊精原细胞的方法,采用0.3%的胰蛋白酶消化新鲜采集的4～6月龄未成年绵羊睾丸组织,制备单细胞悬浮液,经Percoll非连续梯度法离心分离,得到自下而上3条清晰的细胞带,用流式细胞仪对各层细胞进行倍性和抗体染色鉴定.结果表明:第一层细胞中的精原细胞含量达到了76.76%,形态良好,活率高于96%.表明试验中所采用的分离方法对未成年绵羊睾丸精原细胞的分离效果较好,所得细胞可用于进一步的研究.To explore a simple and practical method of separating spermatogonia cells from immature sheep testis,single cell suspension was obtained by digesting freshly collected immature sheep testis with 0.3% trpsin,and subjected to discontinuous Percoll density centrifugation.Three layers of cells were obtained from the bottom to the top.The results showed that the first cell layer contained up to 76.76% spermatogonia with good morphology and viability（above 96%） according to their ploidy analysis and protein expression pattern by flow cytometry.Which indicate that the method used in this study had a good effect in isolating spermatogonia cells from immature sheep testis,and the cells collected could be used for further study.

关 键 词：绵羊 精原细胞 Percoll非连续梯度法 

分 类 号：S826.3[农业科学—畜牧学]