Tissue collection and cell preparation

Primary human epithelial cells were isolated from radical prostatectomy tissues from prostate cancer patients at the University of Illinois at Chicago Medical Center according to guidelines and approval by the Institutional Review Board with written informed consent obtained from all patients. Fresh tissue from the peripheral zone was selected and excised with a 5 mm punch by a pathologist. Final pathology of the tissue was determined by H&E on a thin slice of the punch. The area must be 100% benign to have that classification. Isolation of prostate epithelial cells was as previously described [17, 18] based on the method developed by Donna Peehl [19]. Briefly, tissues were digested with collagenase and plated on collagen-coated dishes in PrEGM (Lonza, Walkersville, MD) for epithelial cell (PrE) growth. Cell type was validated by qRT-PCR for the expression of known basal epithelial cell markers (CK5+, p63+, AR-).