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Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures - a 2 minute Preview of the Experimental Protocol
Wen-Yang Hu, Dan-Ping Hu, Lishi Xie, Lynn A. Birch, Gail S. Prins
University of Illinois at Chicago, Department of Urology, College of Medicine; University of Illinois at Chicago, Department of Pathology, College of Medicine; University of Illinois at Chicago, University of Illinois Cancer Center, College of Medicine;
Using human primary prostate epithelial cells, we report a novel biomarker-free method of functional characterization of stem-like cells by a spheroid-based label-retention assay. A step-by-step protocol is described for BrdU, CFSE, or Far Red 2D cell labeling; three-dimensional spheroid formation; label-retaining stem-like cell identification by immunocytochemistry; and isolation by FACS.
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