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 β and the dorsolateral prostatic secretory protein (mDLP) are given.