Isolation and cultivation of stem cells from adult mouse testes.
The successful isolation and cultivation of spermatogonial stem cells (SSCs) as well as induction of SSCs into pluripotent stem cells will allow us to study their biological characteristics and their applications in therapeutic approaches. Here we provide step-by-step procedures on the basis of previous work in our laboratory for: the isolation of testicular cells from adolescent mice by a modified enzymatic procedure; the enrichment of undifferentiated spermatogonia by laminin selection or genetic selection using Stra8-EGFP (enhanced green fluorescent protein) transgenic mice; the cultivation and conversion of undifferentiated spermatogonia into embryonic stem-like cells, so-called multipotent adult germline stem cells (maGSCs); and characterization of these cells. Normally, it will take about 16 weeks to obtain stable maGSC lines starting from the isolation of testicular cells.
Guan
Kaomei
K
Department of Cardiology and Pneumology, Heart Center, Georg-August-University of Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany. kguan@med.uni-goettingen.de
Wolf
Frieder
F
Becker
Alexander
A
Engel
Wolfgang
W
Nayernia
Karim
K
Hasenfuss
Gerd
G
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049452:Green Fluorescent Proteins; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D013093:Spermatogonia / Q000166:cytology; D013234:Stem Cells / Q000166:cytology*; D013737:Testis / Q000166:cytology*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins