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Clonal Analysis of Embryonic Hematopoietic Stem Cell Precursors Using Single Cell Index Sorting Combined with Endothelial Cell Niche Co-culture
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Biologie du développement
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Journal JoVE Biologie du développement
Clonal Analysis of Embryonic Hematopoietic Stem Cell Precursors Using Single Cell Index Sorting Combined with Endothelial Cell Niche Co-culture

Clonal Analysis of Embryonic Hematopoietic Stem Cell Precursors Using Single Cell Index Sorting Combined with Endothelial Cell Niche Co-culture

DOI:
10.3791/56973-v

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09:32 min

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May 08, 2018

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, , , ,
1Clinical Research Division,Fred Hutchinson Cancer Research Center, 2Department of Pediatrics, Division of Pediatric Hematology/Oncology,University of Washington School of Medicine

Chapitres

  • 00:05Titre
  • 00:52Murine Embryonic Cell Staining
  • 02:49Single Hemogenic Precursor Index Sorting
  • 04:44Clonal Hematopoietic Progeny and Individual Clone Engraftment Property Analyses
  • 07:02Results: Representative Embryonic Hematopoietic Stem Cell Precursor Analysis
  • 09:08Conclusion

Summary

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Here we present methodology for the clonal analysis of hematopoietic stem cell precursors during murine embryonic development. We combine index sorting of single cells from the embryonic aorta-gonad-mesonephros region with endothelial cell co-culture and transplantation to characterize the phenotypic properties and engraftment potential of single hematopoietic precursors.

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Keywords: Clonal AnalysisEmbryonic Hematopoietic Stem Cell PrecursorsSingle Cell Index SortingEndothelial Cell Niche Co-cultureDevelopmental HematopoiesisLineage ContributionsHemogenic PrecursorsPhenotypic PropertiesHSC PrecursorsCollagenaseAntibody CocktailFlow CytometrySingle Cell SortingIndex Sorting

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