流式细胞仪富集小鼠CD90.2阳性精原干细胞

Enrichment of mice spermatogonial stem cells with CD90.2 positive by flow cytometry

目的寻找分离和富集小鼠精原干细胞有效方法。方法首先使用胰酶消化幼鼠的睾丸制成单细胞悬液,联合应用30%percoll液密度梯度离心分离和流式细胞仪分选有CD90.2阳性同时CD117阴性的精原干细胞,并在体外进行原代培养尝试。结果联合应用两种方法获得精原干细胞的纯度高达80.4%,满足体外培养和对其进一步研究要求。结论联合应用两种方法是获得高纯度精原干细胞的有效方法,为精原干细胞培养和研究奠定基础。

Objective To find the effective approach for isolation and enrichment of spermatogonial stem cells. Methods The pup testis were digested with 0.25% trypsin-EDTA into single cell suspension, and then spermatogonial stem cells with CD90.2（＋） and CD117（-） were isolated and enriched by 30% percoll density gradient centrifugation and fluorescent activated cell sorting.the isolated cells were used for primary culture in vitro. Results The number of sorted spermatogonial stem cells by two combined methods accounted for 80.4% of total cell and can be used for in vitro. Conclusion It＇s possible to gain the high purity of spermatogonial stem cells by the combination of 30%percoll and FACS.