Defined Conditions for the Isolation and Expansion of Basal Prostate Progenitor Cells of Mouse and Human Origin

Abstract

SummaryMethods to isolate and culture primary prostate epithelial stem/progenitor cells (PESCs) have proven difficult and ineffective. Here, we present a method to grow and expand both murine and human basal PESCs long term in serum- and feeder-free conditions. The method enriches for adherent mouse basal PESCs with a Lin−SCA-1+CD49f+TROP2high phenotype. Progesterone and sodium selenite are additionally required for the growth of human Lin−CD49f+TROP2high PESCs. The gene-expression profiles of expanded basal PESCs show similarities to ESCs, and NF-kB function is critical for epithelial differentiation of sphere-cultured PESCs. When transplanted in combination with urogenital sinus mesenchyme, expanded mouse and human PESCs generate ectopic prostatic tubules, demonstrating their stem cell activity in vivo. This novel method will facilitate the molecular, genomic, and functional characterization of normal and pathologic prostate glands of mouse and human origin.

Isolation and analysis of discreet human prostate cellular populations

“…Per previous protocols (Goldstein et al, 2011, Hofner et al, 2015), we began by digesting with 1mg/ml collagenase I, II or IV overnight. Crude collagenases, especially type I and II, contain both variable collagenase activity and variable amounts of other enzymes active in the digestive process.…”

“…Furthermore, stem cell surface marker profiles are distinctly found in the basal cells of human prostate epithelium while in mice they are also expressed in a subset of luminal epithelia (Leong et al, 2008, Missol-Kolka et al, 2011). The ex vivo culture conditions for mouse versus human epithelial cells also reveals different nutritional requirements (Hofner et al, 2015, Karthaus et al, 2014). Finally, rats can be induced by various means, notably by hormonal carcinogens to undergo malignant transformation in the prostate (Noble, 1977, Wang and Wong, 1998) while the mouse prostate is relatively resistant to malignant transformation (Shappell et al, 2003).…”

The influence of prostatic anatomy and neurotrophins on basal prostate epithelial progenitor cells

“…This enabled us to investigate their cell surface protein profiles by FACS Array. This screen validated Trop2 and Sca‐1 as being expressed by murine basal PESCs (data not shown, detailed in ). Further, the protein screen identified Syndecan‐1 (CD138) as well as stage specific embryonal antigen ‐ 1 (SSEA‐1) as being expressed on murine basal PESCs, as described in .…”

Protein profile of basal prostate epithelial progenitor cells—stage‐specific embryonal antigen 4 expressing cells have enhanced regenerative potential in vivo

“…Sorted populations were transplanted s.c. in nude mice together with E16 UGSM as described 11, P = 0.01.…”

“…Before conducting histological analyses on fixed tissue, we validated direct Venus fluorescence in freshly dissected s.c. grafts under the fluorescence stereomicroscope. Detection of Venus+ in regenerated prostate tissue (proof of regeneration from transplanted PESCs origin) was done by staining s.c. grafts with a monoclonal antibody against GFP/Venus (ab 290; Abcam, Cambridge, UK) 11.…”