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2. Rescue and Isolation of Rb-deficient Prostate Epithelium by Tissue Recombination
Abstract
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.
Affiliation(s): (2) Department of Urologic Surgery, Department of Cancer Biology, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt Prostate Cancer Center, Vanderbilt University Medical Center, Nashville, TN
(3) Department of Cancer Endocrinology, BC Cancer Agency, Vancouver, BC, Canada
(4) Department of Urology, University of Michigan Comprehensive Cancer Center, Ann Arbor, MI
Series: Methods in Molecular Biology  |  Volume: 218  |  Pub. Date: Dec-09-2002  |  Page Range: 17-33  |  DOI: 10.1385/1-59259-356-9:17
Subject:  Cancer Research
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