The ability to isolate prostate stem cells is essential to explore their role in prostate development and disease. In vitro prostate colonyand sphere-forming assays were used to quantitatively measure murine prostate stem/progenitor cell enrichment and self-renewal. Cell surface markers were screened for their ability to positively or negatively enrich for cells with enhanced growth potential in these assays. Immunohistochemical and FACS analyses demonstrate that specific cell surface markers can be used to discriminate prostate stromal (CD34 ؉ ), luminal epithelial (CD24 ؉ CD49f ؊ ), basal epithelial (CD24 ؉ CD49f ؉ ), hematopoietic (CD45 ؉ , Ter119 ؉ ), and endothelial (CD31 ؉ ) lineages. Sorting for cells with a CD45 ؊ CD31 ؊ Ter119 ؊ Sca-1 ؉ CD49f ؉ antigenic profile results in a 60-fold enrichment for colony-and sphere-forming cells. These cells can self-renew and expand to form spheres for many generations and can differentiate to produce prostatic tubule structures containing both basal and luminal cells in vivo. These cells also localize to the basal cell layer within the region of the gland that is proximal to the urethra, which has been identified as the prostate stem cell niche. Prostate stem cells can be isolated to a purity of up to 1 in 35 by using this antigenic profile. The remarkable similarity in cell surface profile between prostate and mammary gland stem cells suggests these markers may be conserved among epithelial stem cell populations.CD49f ͉ integrin ␣6 ͉ Sca-1 ͉ CD24 heat-stable antigen ͉ stem cell niche S tem cells are of interest clinically because of their potential to repair damaged tissues, treat degenerative diseases, and because of their purported role in tumor initiation. The ability to identify and isolate stem cells is necessary to study their specialized biology. Enrichment for many types of tissue stem cells has been achieved by using cell surface markers. Murine hematopoietic stem cells can be enriched by sorting Lin Ϫ Thy-1 lo Sca-1 ϩ ckit ϩ cells from the bone marrow (1). Recent studies suggest that even better purity can be achieved by further sorting based on expression of the SLAM family receptors CD150 and CD48 (2). Bronchioavelolar stem cells can be isolated from their niche at the bronchioalveolar duct junction (BADJ) by sorting cells with a CD45 Ϫ CD31 Ϫ Sca-1 ϩ CD34 ϩ profile (3). Data from two recent reports show that mouse mammary stem cells possess a Lin Ϫ Sca-1 ϩ CD140a Ϫ CD24 ϩ CD49f ϩ CD29 ϩ cell surface profile and can be isolated to a purity of up to 1 in 20 by using subsets of these markers (4, 5).The presence of stem cells in the prostate first was proposed to explain the seemingly inexhaustible capacity of the organ to regenerate during androgen cycling experiments (6). The identification of side-population cells and replication quiescent BrdU label-retaining cells further suggests that stem cells exist in the gland (7,8). Several studies have enriched for primitive prostate cells by using cell surface markers. Richardson et al. (9) demonstrated that th...

The epithelium of the adult prostate contains 3 distinct cell types: basal, luminal, and neuroendocrine. Tissue-regenerative activity has been identified predominantly from the basal cells, isolated by expression of CD49f and stem cell antigen-1 (Sca-1). An important question for the field is whether all basal cells have stem cell characteristics. Prostate-specific microarray databases were interrogated to find candidate surface antigens that could subfractionate the basal cell population. Tumor-associated calcium signal transducer 2 (TACSTD2/Trop2/M1S1/GA733-1) was identified because it was enriched after castration, in prostate sphere cells and in the basal fraction. In the murine prostate, Trop2 shows progenitor characteristics such as localization to the region of the gland proximal to the urethra and enrichment for sphere-forming and colony-forming cells. Trop2 subfractionates the basal cells into 2 populations, both of which express characteristic basal cell markers by quantitative PCR. However, only the basal cells expressing high levels of Trop2 were able to efficiently form spheres in vitro. In the human prostate, where Sca-1 is not expressed, sphere-forming progenitor cells were also isolated based on high expression of Trop2 and CD49f. Trop2-expressing murine basal cells could regenerate prostatic tubules in vivo, whereas the remaining basal cells had minimal activity. Evidence was found for basal, luminal, and neuroendocrine cells in prostatic tubules regenerated from Trop2 hi basal cells. In summary, functionally distinct populations of cells exist within the prostate basal compartment and an epithelial progenitor can give rise to neuroendocrine cells in vivo.neuroendocrine ͉ progenitor ͉ sphere assay

SUMMARY MYCN amplification and overexpression are common in neuroendocrine prostate cancer (NEPC). However, the impact of aberrant N-Myc expression in prostate tumorigenesis and the cellular origin of NEPC have not been established. We define N-Myc and activated AKT1 as oncogenic components sufficient to transform human prostate epithelial cells to prostate adenocarcinoma and NEPC with phenotypic and molecular features of aggressive, late-stage human disease. We directly show that prostate adenocarcinoma and NEPC can arise from a common epithelial clone. Further, N-Myc is required for tumor maintenance and destabilization of N-Myc through Aurora A kinase inhibition reduces tumor burden. Our findings establish N-Myc as a driver of NEPC and a target for therapeutic intervention.