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Capítulos
- 00:04Título
- 01:16Culture and Maintenance of HPrEC Cells in 2D
- 02:30Labeling HPrEC with BrdU, CFSE, or Far Red Pro-dyes
- 03:24Prostasphere Formation in the 3D Basement Membrane Culture System
- 04:27Isolation of the CFSE Label-retaining Prostate Stem Cells by FACS Sorting
- 06:45Results: Evaluation of the Sphere Based Label-retaining Assay
- 08:03Conclusion
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.
