Murine spermatogonial stem cells: targeted transgene expression and purification in an active state
Murine spermatogonial stem cells: targeted transgene expression and purification in an active state
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DOI：
10.1093/embo-reports/kvf149
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发表时间：
2002-08-01
期刊：
影响因子：
7.7
通讯作者：
Cuzin, F
中科院分区：
文献类型：
--
作者：
Giuili, G;Tomljenovic, A;Cuzin, F
精原细胞特异性 Stra8 位点的 400 bp 片段足以将基因表达引导至转基因小鼠的生殖干细胞。基于细胞的免疫磁性分选设计了分级分离程序，其中启动子驱动表面功能中性蛋白标签的表达。纯化的细胞表达已知的精原细胞 Rbm、细胞周期蛋白 A2 和 EP-Cam 分子标记，以及干细胞组分特有的 β(1) 和 α(6) 整合素。干细胞富集了 700 倍，这是通过纯化的级分在生殖细胞耗尽的受体睾丸中重建精子发生的能力来确定的。
A 400 bp fragment of the spermatogonia-specific Stra8 locus was sufficient to direct gene expression to the germinal stem cells in transgenic mice. A fractionation procedure was devised, based on immunomagnetic sorting of cells in which the promoter drives the expression of a surface functionally neutral protein tag. The purified cells expressed the known molecular markers of spermatogonia Rbm, cyclin A2 and EP-Cam, and the beta(1)- and alpha(6)-integrins characteristic of the stem cell fraction. A 700-fold enrichment in stem cells was determined by the ability of the purified fractions to re-establish spermatogenesis in germ cell-depleted recipient testes.