Isolation, cultivation and characterization of adult murine prostate stem cells

Lukacs, Rita U.; Goldstein, Andrew S.; Lawson, Devon A.; Cheng, Donghui; Witte, Owen N.

doi:10.1038/nprot.2010.11

Cited by 175 publications

(228 citation statements)

References 18 publications

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“…Most importantly, the finding that cells of origin of prostate cancer reside in the basal compartment is consistent with findings in human and murine tissues by Witte and coinvestigators who used high expression of Trop2 and CD49f to enrich for prostatic basal cells [3,4,33]. In addition to this, Wang et al showed luminal stem cells (CARNs) are also a cell of origin of prostate cancer in mouse prostate [1].…”

Section: Discussionsupporting

confidence: 71%

Human Epithelial Basal Cells Are Cells of Origin of Prostate Cancer, Independent of CD133 Status

et al. 2012

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Normal prostatic epithelium is composed of basal and luminal cells. Prostate cancer can be initiated in both benign basal and luminal stem cells, but because basal cell markers are not expressed in patient tumors, the former result was unexpected. Since the cells of origin of prostate cancer are important therapeutic targets, we sought to provide further proof that basal stem cells have tumorigenic potential. Prostatic basal cells were enriched based on a2b1integrin hi expression and further enriched for stem cells using CD133 in nontumorigenic BPH-1 cells. Human embryonic stem cells (hESCs) were also used as a source of normal stem cells. To test their tumorigenicity, we used two alternate stromal-based approaches; (a) recombination with human cancer-associated fibroblasts (CAFs) or (b) recombination with embryonic stroma (urogenital mesenchyme) and treated host mice with testosterone and 17b-estradiol. Enriched a2b1integrin hi basal cells from BPH-1 cells resulted in malignant tumor formation using both assays of tumorigenicity. Surprisingly, the tumorigenic potential did not reside in the CD133 1 stem cells but was consistently observed in the CD133 2 population. CAFs also failed to induce prostatic tumors from hESCs. These data confirmed that benign human basal cells include cells of origin of prostate cancer and reinforced their importance as therapeutic targets. In addition, our data suggested that the more proliferative CD133 2 basal cells are more susceptible to tumorigenesis compared to the CD133 1 -enriched stem cells. These findings challenge the current dogma that normal stem cells and cells of origin of cancer are the same cell type(s).

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Section: Discussionsupporting

confidence: 71%

Human Epithelial Basal Cells Are Cells of Origin of Prostate Cancer, Independent of CD133 Status

et al. 2012

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“…Before performing the coinjections, UGSM was prepared freshly from foetuses of E16 C57Bl/6 mice as previously described by Lukacs et al . 6. Briefly, timed pregnancies of C57/BL6 mice were set up and pregnant females were killed at day 16 (E16).…”

Section: Methodsmentioning

confidence: 99%

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

Höfner

Klein

Eisen

et al. 2016

J Cellular Molecular Medi

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The long‐term propagation of basal prostate progenitor cells ex vivo has been very difficult in the past. The development of novel methods to expand prostate progenitor cells in vitro allows determining their cell surface phenotype in greater detail. Mouse (Lin−Sca‐1+ CD49f+ Trop2high‐phenotype) and human (Lin− CD49f+ TROP2high) basal prostate progenitor cells were expanded in vitro. Human and mouse cells were screened using 242 anti‐human or 176 antimouse monoclonal antibodies recognizing the cell surface protein profile. Quantitative expression was evaluated at the single‐cell level using flow cytometry. Differentially expressed cell surface proteins were evaluated in conjunction with the known CD49f+/TROP2high phenotype of basal prostate progenitor cells and characterized by in vivo sandwich‐transplantation experiments using nude mice. The phenotype of basal prostate progenitor cells was determined as CD9+/CD24+/CD29+/CD44+/CD47+/CD49f+/CD104+/CD147+/CD326+/Trop2high of mouse as well as human origin. Our analysis revealed several proteins, such as CD13, Syndecan‐1 and stage‐specific embryonal antigens (SSEAs), as being differentially expressed on murine and human CD49f+ TROP2+ basal prostate progenitor cells. Transplantation experiments suggest that CD49f+ TROP2high SSEA‐4high human prostate basal progenitor cells to be more potent to regenerate prostate tubules in vivo as compared with CD49f+ TROP2high or CD49f+ TROP2high SSEA‐4low cells. Determination of the cell surface protein profile of functionally defined murine and human basal prostate progenitor cells reveals differentially expressed proteins that may change the potency and regenerative function of epithelial progenitor cells within the prostate. SSEA‐4 is a candidate cell surface marker that putatively enables a more accurate identification of the basal PESC lineage.

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“…CK8 expression was not observed in WT and Abcg2 null spheres at day 14 using immunofluorescence staining. Previous studies document inability to detect high CK8 protein expression in spheres before day 14 [31]. Dual immunofluorescence staining of p63 and CK5 showed heterogeneity in terms of colocalization of p63 and CK5.…”

Section: Fig 3 Sphere Formation Efficiency Is Increased With Enhancmentioning

confidence: 99%

“…2B). The sphere formation efficiency determines in vitro proliferation and differentiation potential of basal and stem/progenitor cells [31]. Although the number of p63 expressing basal cells was not changed after one and five cycles of regression and regeneration (Fig.…”

Section: Luminal Cell Number Is Decreased In Abcg2 Null Ventral Prostmentioning

confidence: 99%

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Multi-Drug Resistance ABC Transporter Inhibition Enhances Murine Ventral Prostate Stem/Progenitor Cell Differentiation

Samant

Jackson

Felix

et al. 2015

Stem Cells and Development

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Multi-drug resistance (MDR)-ATP binding cassette (ABC) transporters, ABCB1, ABCC1, and ABCG2 participate in the efflux of steroid hormones, estrogens, and androgens, which regulate prostate development and differentiation. The role of MDR-ABC efflux transporters in prostate epithelial proliferation and differentiation remains unclear. We hypothesized that MDR-ABC transporters regulate prostate differentiation and epithelium regeneration. Prostate epithelial differentiation was studied using histology, sphere formation assay, and prostate regeneration induced by cycles of repeated androgen withdrawal and replacement. Embryonic deletion of Abcg2 resulted in a decreased number of luminal cells in the prostate and increased sphere formation efficiency, indicating an imbalance in the prostate epithelial differentiation pattern. Decreased luminal cell number in the Abcg2 null prostate implies reduced differentiation. Enhanced sphere formation efficiency in Abcg2 null prostate cells implies activation of the stem/progenitor cells. Prostate regeneration was associated with profound activation of the stem/progenitor cells, indicating the role of Abcg2 in maintaining stem/progenitor cell pool. Since embryonic deletion of Abcg2 may result in compensation by other ABC transporters, pharmacological inhibition of MDR-ABC efflux was performed. Pharmacological inhibition of MDR-ABC efflux enhanced prostate epithelial differentiation in sphere culture and during prostate regeneration. In conclusion, Abcg2 deletion leads to activation of the stem/progenitor cells and enhances differentiating divisions; and pharmacological inhibition of MDR-ABC efflux leads to epithelial differentiation. Our study demonstrates for the first time that MDR-ABC efflux transporter inhibition results in enhanced prostate epithelial cell differentiation.

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Isolation, cultivation and characterization of adult murine prostate stem cells

Cited by 175 publications

References 18 publications

Human Epithelial Basal Cells Are Cells of Origin of Prostate Cancer, Independent of CD133 Status

Human Epithelial Basal Cells Are Cells of Origin of Prostate Cancer, Independent of CD133 Status

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

Multi-Drug Resistance ABC Transporter Inhibition Enhances Murine Ventral Prostate Stem/Progenitor Cell Differentiation

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