Isolation and functional characterization of murine prostate stem cells
Abstract
The ability to isolate prostate stem cells is essential to explore their role in prostate development and disease. In vitro prostate colony- and 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.
Acknowledgments
We thank Connie Eaves, Robert Signer, Encarnacion Montecino-Rodriguez, and Caius Radu for advice on project design and FACS sorting and analysis. This work was supported by funds from the Prostate Cancer Foundation and the University of California, Los Angeles Special Program of Research Excellence in Prostate Cancer, (Jean deKernion, principal investigator). O.N.W. is an Investigator of the Howard Hughes Medical Institute. D.A.L. is supported by National Institutes of Health/Public Health Services Grant T32 CA09056 Tumor Cell Biology Training Grant.
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© 2006 by The National Academy of Sciences of the USA. Freely available online through the PNAS open access option.
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Received: October 19, 2006
Published online: January 2, 2007
Published in issue: January 2, 2007
Keywords
Acknowledgments
We thank Connie Eaves, Robert Signer, Encarnacion Montecino-Rodriguez, and Caius Radu for advice on project design and FACS sorting and analysis. This work was supported by funds from the Prostate Cancer Foundation and the University of California, Los Angeles Special Program of Research Excellence in Prostate Cancer, (Jean deKernion, principal investigator). O.N.W. is an Investigator of the Howard Hughes Medical Institute. D.A.L. is supported by National Institutes of Health/Public Health Services Grant T32 CA09056 Tumor Cell Biology Training Grant.
Notes
This article contains supporting information online at www.pnas.org/cgi/content/full/0609684104/DC1.
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The authors declare no conflict of interest.
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Isolation and functional characterization of murine prostate stem cells, Proc. Natl. Acad. Sci. U.S.A.
104 (1) 181-186,
https://doi.org/10.1073/pnas.0609684104
(2007).
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