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Chapters
- 00:04Title
- 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
Ved hjælp af humane primære prostata epiteliale celler, rapporterer vi en roman biomarkør-fri metode til funktionel karakterisering af Stem-lignende celler ved en spheroid-baseret etiket-retention assay. En trin-for-trin-protokol er beskrevet for BrdU, CFSE eller langt rødt 2D-celle mærkning; tredimensionel sfærisk dannelse; etiket-fastholde Stem-lignende celle identifikation ved immunocytochemistry; og isolation af FACS.
