Technical Reference #1790

1790.

Capacity for stochastic self-renewal and differentiation in mammalian spermatogonial stem cells Zhuoru Wu; Katherine Luby-Phelps; Abhijit Bugde; Laura A. Molyneaux; Bray Denard; Wen-Hong Li; Guerol M. Sueel; David L. Garbers, University of Texas Southwestern Medical Center, Journal of Cell Biology, 187(1790), (2009)
Link To Paper

Materials & Methods:
Cell culture media PBS nonessential amino acids MEM vitamin solution l-glutamine solution 0.05% or 0.25% trypsin-EDTA solutions B27 minus vitamin A supplement antibiotic-antimycotic solutions and horse serum were obtained from Invitrogen. BSA and DMSO were obtained from EMD. Mitomycin-C mouse laminin mouse EGF rat GDNF bovine apo-transferrin human basic fibroblast growth factor (bFGF) d-(+)-glucose d-biotin ascorbic acid sodium pyruvate putrescine progesterone -estradiol 17-cypionate insulin sodium selenite 2-mercaptoethanol (ME) dl-lactic acid and all fatty acids were obtained from Sigma-Aldrich. Mouse leukemia inhibitory factor/ESGRO (LIF) was obtained from Millipore. FBS was obtained from Atlanta Biologicals. Dispase and rat-tail collagen I-coated culture dishes were obtained from Thermo Fisher Scientific Inc. The In Situ Cell Death Detection kit (TMR red) was obtained from Roche. Busulfan was obtained from MP Biomedicals. Glass-bottom dishes were obtained from MatTek Corporation.

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
SSC; spermatogonial stem cell.