Murine spermatogonial stem cells: targeted transgene expression and purification in an active state.
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.
Giuili
Galicia
G
Unité 470 de l'INSERM, Université de Nice, France.
Tomljenovic
Andrea
A
Labrecque
Nathalie
N
Oulad-Abdelghani
Mustapha
M
Rassoulzadegan
Minoo
M
Cuzin
François
F
D000367:Age Factors; D000818:Animals; D001483:Base Sequence; D015704:CD4 Antigens / Q000096:biosynthesis; D002469:Cell Separation / Q000379:methods*; D019925:Cyclin A / Q000378:metabolism; D056751:Cyclin A2; D018076:DNA, Complementary / Q000378:metabolism; D005091:Exons; D015723:Gene Library; D017930:Genes, Reporter; D006801:Humans; D007150:Immunohistochemistry; D039503:Integrin alpha6 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008280:Magnetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010957:Plasmids; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000378:metabolism*; D013737:Testis / Q000378:metabolism / Q000473:pathology; D014018:Tissue Distribution; D019076:Transgenes
C503721:CCNA2 protein, human; D015704:CD4 Antigens; D019925:Cyclin A; D056751:Cyclin A2; D018076:DNA, Complementary; D039503:Integrin alpha6; D019012:Integrin beta1; D009687:Nuclear Proteins; C501499:RBMY1A1 protein, human; D016601:RNA-Binding Proteins; C501500:Rbmy protein, mouse; D008156:Luciferases