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Isolation of Embryonic Hematopoietic Niche Cells by Flow Cytometry and Laser Capture Microdissection

  • Daisuke Sugiyama
  • Tatsuya Sasaki
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1035)

Abstract

Hematopoietic stem cells (HSCs) can differentiate into several types of hematopoietic cells, such as erythrocytes, megakaryocytes, lymphocytes, neutrophils, or macrophages, and also undergo self-renewal to sustain hematopoiesis throughout an organism’s lifetime. HSCs emerge and expand during mouse embryogenesis. HSC regulation is governed by two types of activity: intrinsic activity programmed primarily by cell autonomous gene expression, and extrinsic factors, which originate from the so-called niche cells surrounding HSCs. Previously, we reported that endothelial niche cells regulate HSC generation at aorta-gonad-mesonephros region and placenta, and that hepatoblastic niche cells regulate HSC differentiation in mouse embryonic liver. In the course of those studies, we employed immunohistochemistry, flow cytometry, and the laser capture microdissection system to assess embryonic regulation of the mouse hematopoietic niche.

Key words

Hematopoietic stem cells Niche cells Embryo Visualization Laser capture microdissection 

Notes

Acknowledgments

We thank The Ministry of Education, Culture, Sports, Science and Technology; The Ministry of Health, Labor and Welfare; and The Japan Society for the Promotion of Science for grant support, and Dr. Elise Lamar for critical reading of the manuscript.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Daisuke Sugiyama
    • 1
  • Tatsuya Sasaki
    • 1
  1. 1.Division of Hematopoietic Stem Cells, Advanced Medical Initiatives, Department of Advanced Medical InitiativesKyushu University Faculty of Medical SciencesFukuokaJapan

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