In vitro propagation and characterization of neoplastic stem/progenitor-like cells from human prostate cancer tissue
Article first published online: 30 JUL 2009
Copyright © 2009 Wiley-Liss, Inc.
Volume 69, Issue 15, pages 1683–1693, 1 November 2009
How to Cite
Guzmán-Ramírez, N., Völler, M., Wetterwald, A., Germann, M., Cross, N. A., Rentsch, C. A., Schalken, J., Thalmann, G. N. and Cecchini, M. G. (2009), In vitro propagation and characterization of neoplastic stem/progenitor-like cells from human prostate cancer tissue. Prostate, 69: 1683–1693. doi: 10.1002/pros.21018
- Issue published online: 20 SEP 2009
- Article first published online: 30 JUL 2009
- Manuscript Accepted: 24 JUN 2009
- Manuscript Received: 27 FEB 2009
- 6th Framework Program of the European Community. Grant Number: PROMET-018858
- Department of Clinical Research, University of Bern
- cancer stem/progenitor-like cells;
According to the cancer stem cell hypothesis, tumor growth is sustained by a subpopulation of cancer stem/progenitor-like cells. Self-renewal and high clonogenic potential are characteristics shared by normal stem and neoplastic stem/progenitor-like cells. We investigated whether human prostate cancer specimens contain cells with these properties.
Self-renewal and clonogenic potential were assessed by serial passaging of spheres and colony formation, respectively. Gene expression was analyzed by real time PCR. Protein expression was detected by immunocytochemistry. The neoplastic nature of the cells was verified by detection of the TMPRSS2/ERG gene fusion expression.
The epithelial fraction isolated from surgical specimens generated colonies in 68% (19/28) of the patients. Laminin adhesion selected for cells with high clonogenic potential. The epithelial fraction from 85% (42/49) of the patients generated primary prostaspheres. Serial passaging of prostaspheres demonstrated their self-renewal capacity, which is also supported by their expression of the stem cell markers Oct-4, Nanog, Bmi-1, and Jagged-1 mRNA. Cells derived from prostaspheres were more clonogenic than the parental epithelial fraction. The pattern of mRNA expression in prostaspheres resembled that of the basal compartment of the prostate (CK5+/CK14+/CK19high/CK18−/low/c-met+/AR−/low/PSA−/low), but also included stem cell markers (CD49b+/CD49f+/CD44+/ΔNp63+/Nestin+/CD133+). The distribution of marker expression in prostaspheres suggests their heterogeneous cell composition. Prostaspheres expressed significantly higher PSCA mRNA levels than the epithelial fraction.
Human prostate cancer specimens contain neoplastic cells with self-renewal and clonogenic potential, which can be enriched and perpetuated in prostaspheres. Prostaspheres should prove valuable for the identification of prostate cancer stem/progenitor-like cells. Prostate 69: 1683–1693, 2009. © 2009 Wiley-Liss, Inc.