Rescue and isolation of Rb-deficient prostate epithelium by tissue recombination.
The ability to rescue viable prostate precursor tissue from Rb-/-fetal mice has allowed for the generation of Rb-/-prostate tissue and Rb-/-prostate epithelial cell lines. Herein, we provide a protocol for the rescue of urogenital precursor tissue from mouse embryos harboring the lethal Rb-/-mutation. The rescued precursors can matured as subrenal capsule grafts in athymic mice. Subsequently prostatic tissue can be used as a source for Rb-/-epithelium in a tissue recombination protocol for the generation of chimeric prostate grafts in athymic male mouse hosts. We have also provided a detailed description for isolating and propagating the Rb-/-epithelium from such tissue recombinants as established cell lines. Methods for characterizing the grafts and cell lines by determining the retention of prostate-specific epithelial expression markers, including cytokeratins, the androgen receptor, estrogen receptor beta and the dorsolateral prostatic secretory protein (mDLP) are given.
Hayward
Simon W
SW
Department of Urologic Surgery, Department of Cancer Biology, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt Prostate Cancer Center, Vanderbilt University Medical Center, Nashville, TN, USA.
Wang
Yuzhuo
Y
Day
Mark L
ML
D000818:Animals; D002460:Cell Line; D002678:Chimera; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008819:Mice, Nude; D008967:Molecular Biology / Q000379:methods*; D011247:Pregnancy; D011467:Prostate / Q000166:cytology*; D011471:Prostatic Neoplasms; D051381:Rats; D017207:Rats, Sprague-Dawley; D011995:Recombination, Genetic; D016160:Retinoblastoma Protein / Q000235:genetics*; D013234:Stem Cells / Q000166:cytology
D016160:Retinoblastoma Protein