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Identification and Isolation of Burst-Forming Unit and Colony-Forming Unit Erythroid Progenitors from Mouse Tissue by Flow Cytometry
Journal JoVE
Biologie du développement
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Journal JoVE Biologie du développement
Identification and Isolation of Burst-Forming Unit and Colony-Forming Unit Erythroid Progenitors from Mouse Tissue by Flow Cytometry
DOI:

08:31 min

November 04, 2022

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Chapitres

  • 00:05Introduction
  • 00:47Bone Marrow Cell Extraction
  • 02:07Spleen Cell Extraction
  • 02:46Antibody Staining of FMOs and Single-Color Controls
  • 03:53Cell Resuspension
  • 04:38Sample Preparation for Flow Cytometric Analysis
  • 05:19Gating Strategy for Setting up the Sorting Gates
  • 06:49Results: Effect of Erythropoietin on Bone Marrow and Spleen Progenitors
  • 07:42Conclusion

Summary

Traduction automatique

Here, we describe a novel flow cytometric method for prospective isolation of early burst-forming unit erythroid (BFU-e) and colony-forming unit erythroid (CFU-e) progenitors directly from fresh mouse bone marrow and spleen. This protocol, developed based on single-cell transcriptomic data, is the first to isolate all the tissue's erythroid progenitors with high purity.

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