Murine spermatogonial stem cells: targeted transgene expression and purification in an active state

小鼠精原干细胞:活性状态下的靶向转基因表达和纯化

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作者:Galicia Giuili, Andrea Tomljenovic, Nathalie Labrecque, Mustapha Oulad-Abdelghani, Minoo Rassoulzadegan, François Cuzin

Abstract

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 beta1- and alpha6-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.

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