168 DEFINING CONDITIONS TO GENERATE AND MAINTAIN PROSTATE CANCER STEM CELLS

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2011168 DEFINING CONDITIONS TO GENERATE AND MAINTAIN PROSTATE CANCER STEM CELLS Adrian Rybak, Anil Kapoor, Lizhi He, and Damu Tang Adrian RybakAdrian Rybak Hamilton, Canada More articles by this author , Anil KapoorAnil Kapoor Hamilton, Canada More articles by this author , Lizhi HeLizhi He Hamilton, Canada More articles by this author , and Damu TangDamu Tang Hamilton, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.237AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Cancer stem cells (CSCs) play critical roles in cancer metastasis and chemoresistance. In vitro, CSCs can be cultured in defined serum-free media (SFM) as suspension spheres. While accumulating evidence demonstrates the existence of prostate cancer stem cells (PCSCs), PCSCs have not been isolated from human primary prostate cancer (PC) tissues. This is largely due to limited primary PC tissues available clinically, as well as the heterogeneous nature of primary PC. These limitations have directly hindered the isolation of PCSCs from human primary PC tissues using classical methods that have been utilized to isolate CSCs from brain and breast cancers. These classical methods involved affinity purification of CSCs using specific markers for the respective tissue stem cells. Therefore, finding methods to propagate PCSCs may serve as an alternative means of isolating and enriching for this sub-population of cells from primary PC. To overcome these limitations, we have developed a unique condition to culture stem-like cells from the DU145 PC cell line. METHODS DU145 PC cells were plated under defined serum-free conditions, allowing a sub-population of stem-like cells to propagate as suspended spheroid colonies, or ‘spheres'. Immunofluorescent staining of spheres was carried out to identify potential cell surface markers, and to aid in the identification of the prospective tumor cell of origin. Furthermore, subcutaneous implantations were carried out in NOD/SCID mice in order to study the in vivo tumorigenic properties of sphere cells. RESULTS A sub-population of PC stem-like cells have been isolated and cultured long-term as spheres without any apparent reduction in their stem cell-like properties. These cells express prostate basal cell markers (CD44+34βE12+integrin-α2β1+) and have been shown to initiate tumorigenesis when implanted in NOD/SCID mice. Consistent with these reports, sphere cells displayed 100-fold increases in tumor-forming ability compared to monolayer cells in NOD/SCID mice, as 103, 104, and 105 sphere cells generated xenograft tumors as efficient as 105, 106, and 107 monolayer cells, respectively. Furthermore, stem-like cells can be re-isolated from xenograft tumors using our culture conditions. CONCLUSIONS We have defined culture conditions for long-term maintenance of sphere-forming cells which display stem-like properties from the DU145 cell line. Our defined serum-free culture conditions may serve as an alternative method to isolating and enriching for PCSCs from primary PC tissues. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e70-e71 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Adrian Rybak Hamilton, Canada More articles by this author Anil Kapoor Hamilton, Canada More articles by this author Lizhi He Hamilton, Canada More articles by this author Damu Tang Hamilton, Canada More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...