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Murine spermatogonial stem cells: targeted transgene expression and purification in an active state
Author(s) -
Giuili Galicia,
Tomljenovic Andrea,
Labrecque Nathalie,
OuladAbdelghani Mustapha,
Rassoulzadegan Minoo,
Cuzin François
Publication year - 2002
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1093/embo-reports/kvf149
Subject(s) - biology , stem cell , transgene , microbiology and biotechnology , somatic cell , cell sorting , gene , flow cytometry , genetics
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 β 1 ‐ and α 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.