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Improving the Accuracy of Flow Cytometric Assessment of Mitochondrial Membrane Potential in Hematopoietic Stem and Progenitor Cells Through the Inhibition of Efflux Pumps
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Biologia
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JoVE Journal Biologia
Improving the Accuracy of Flow Cytometric Assessment of Mitochondrial Membrane Potential in Hematopoietic Stem and Progenitor Cells Through the Inhibition of Efflux Pumps

Improving the Accuracy of Flow Cytometric Assessment of Mitochondrial Membrane Potential in Hematopoietic Stem and Progenitor Cells Through the Inhibition of Efflux Pumps

DOI:
10.3791/60057-v

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07:17 min

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July 30, 2019

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, ,
1Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research,Albert Einstein College of Medicine, 2Departments of Cell Biology and Stem Cell Institute,Albert Einstein College of Medicine, 3Department of Medicine,Albert Einstein College of Medicine, 4Albert Einstein Cancer Center and Diabetes Research Center,Albert Einstein College of Medicine

Capitoli

  • 00:04Titolo
  • 00:43Bone Marrow Isolation
  • 02:22Immunostaining for Detection of Hematopoietic Stem Cells
  • 03:28TMRM Staining
  • 04:17Flow Cytometry
  • 05:38Results: Flow Cytometry Analysis of Mitochondrial Membrane Potential
  • 06:33Conclusion

Summary

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Xenobiotic efflux pumps are highly active in hematopoietic stem and progenitor cells (HSPCs) and cause extrusion of TMRM, a mitochondrial membrane potential fluorescent dye. Here, we present a protocol to accurately measure mitochondrial membrane potential in HSPCs by TMRM in the presence of Verapamil, an efflux pump inhibitor.

Tags

Keywords: Flow CytometryMitochondrial Membrane PotentialHematopoietic Stem CellsProgenitor CellsEfflux PumpsTMRMVerapamilBone Marrow IsolationLineage CocktailCD135

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