人睾丸精原细胞的分离和纯化  

Separation and purification of spermatogonia

作　　者：毕罡[1] 李彦锋[1] 李黔生[1] 靳风烁[1] 张勇[1] 徐序广[1] 靳文生[1] 霍文谦[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所泌尿外科,重庆400042

出　　处：《第三军医大学学报》2005年第11期1142-1144,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 2 0 0 0 93)~~

摘　　要：目的　探讨人睾丸精原细胞的分离纯化。方法　用联合二步酶消化法获得人生殖细胞悬液,经Percoll不连续密度梯度离心,再用差异粘附法纯化得到成活率较好、含量较高的人精原细胞。结果　所获得睾丸组织细胞悬液内活细胞、死细胞平均所占百分比分别为89 71%、10 2 9%。Percoll不连续密度梯度中,细胞主要在相邻梯度的界面处形成细胞带,其中精原细胞主要分布于2 7%～3 5 %Percoll梯度间,根据细胞体外培养时的形态学特征等进行判断和分析,经纯化后精原细胞的纯度达到60 42 %。结论　应用联合酶消化法、Percoll不连续密度梯度离心和差异粘附等方法可以有效地对人精原细胞进行分离和纯化。Objective To study separation and purification method of human fetal spermatogonia. Methods Sequential two-step enzymatic digestion was used to prepare germ cell suspension of human fetus, then Percoll discontinue density gradient centrifugation and isolation according to the different adhesiveness were used to purify spermatogonia. Results The average percentages of living cells, dead cells in suspension were 89.71% and 10.29% respectively. Spermatogonia were mainly distributed in Percoll gradient between 27%-35%. Purity of spermatogonia was 60.42% after observing the morphological characteristics of spermatogonia. Conclusion Human fetal spermatogonia can be isolated and purified by sequential two-step enzymatic digestion and Percoll discontinue density gradient centrifugation.

关 键 词：精原细胞 分离 纯化 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学] R322.64[医药卫生—基础医学]