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Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures
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JoVE Journal Cancer Research
Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures

Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures

DOI:
10.3791/60357-v

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09:06 min

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December 13, 2019

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, , , ,
1Department of Urology, College of Medicine,University of Illinois at Chicago, 2Department of Pathology, College of Medicine,University of Illinois at Chicago, 3University of Illinois Cancer Center, College of Medicine,University of Illinois at Chicago

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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: Stem CellsSpheroid CultureLabel-retention AssayCFSEFar RedProstate CancerEpithelial Cells3D CultureCell IsolationTherapeutic ResistanceCancer Stem Cells

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