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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 an effective method to isolate and enrich mouse spermatogonial stem cells. Methods The testis of rat pups was digested with trypsin to make a single cell suspension. Serum CD90.2-positive and CD117-negative spermatogonial stem cells were sorted by liquid chromatography with 30% percoll density gradient centrifugation and flow cytometry. For primary culture try. Results The purity of spermatogonial stem cells obtained by the combination of two methods was up to 80.4%, which met the requirements of in vitro culture and further research. Conclusion The combination of these two methods is an effective method to obtain high purity spermatogonial stem cells and lay a foundation for the culture and research of spermatogonial stem cells.