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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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Developmental Biology
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JoVE Journal Developmental Biology
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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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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