Long-term proliferation and characterization of human spermatogonial stem cells obtained from obstructive and non-obstructive azoospermia under exogenous feeder-free culture conditions
Long-term proliferation and characterization of human spermatogonial stem cells obtained from obstructive and non-obstructive azoospermia under exogenous feeder-free culture conditions
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DOI：
10.1111/j.1365-2184.2010.00691.x
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发表时间：
2010-08-01
影响因子：
8.5
通讯作者：
Lee, D. R.
中科院分区：
文献类型：
--
作者：
Lim, J. J.;Sung, S. -Y.;Lee, D. R.
目的：本研究的目的是提高从阻塞性无精子症（OA）和非阻塞性无精子症（NOA）患者中获得的人精原干细胞（SSC）的分离效率并优化其增殖潜力，并进一步表征这些细胞在不孕症治疗或生殖生物学研究中的潜在用途。材料和方法：我们应用细胞分选方法，利用胶原蛋白和磁激活细胞分离来克服障碍，开发收集系统和简单的长期增殖系统，可从阻塞性 OA 和 NOA 患者中产生大量高纯度 SSC。结果：源自 OA 和 NOA 患者的 SSC 在体外增殖并保持其特性超过 12 代（> 6 个月）。此外，SSC 特异性标记物 GFR α-1 和整合素 α 6 呈阳性的细胞群在第 8 代时增加到 80% 以上。结论：这些发现可能支持这样的观点：SSC 的体外增殖可能是不孕症治疗和生殖生物学研究的有用工具。
Objectives:The aim of the present study was to improve efficiency of isolation and to optimize proliferative potential of human spermatogonial stem cells (SSCs) obtained from obstructive azoospermic (OA) and non-obstructive azoospermic (NOA) patients, and further, to characterize these cells for potential use in infertility treatment or study of reproductive biology.Materials and methods:We have applied a cell-sorting method, using collagen and magnetic activated cell separation to overcome obstacles, developing a collection system, and simple long-term proliferation system, that yields large numbers of high-purity SSCs from obstructive OA and NOA patients.Results:SSCs derived from OA and NOA patients proliferated and maintained their characteristics for more than 12 passages (> 6 months) in vitro. Moreover, the population of cells positive for the SSC-specific markers GFR alpha-1 and integrin alpha 6, increased to more than 80% at passage 8.Conclusion:These finding may support the idea that in vitro propagation of SSCs could be a useful tool for infertility treatment and study of reproductive biology.