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Haematopoietic stem cells derive directly from aortic endothelium during development

Nature volume 464, pages 108–111 (2010)Cite this article

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Abstract

A major goal of regenerative medicine is to instruct formation of multipotent, tissue-specific stem cells from induced pluripotent stem cells (iPSCs) for cell replacement therapies. Generation of haematopoietic stem cells (HSCs) from iPSCs or embryonic stem cells (ESCs) is not currently possible, however, necessitating a better understanding of how HSCs normally arise during embryonic development. We previously showed that haematopoiesis occurs through four distinct waves during zebrafish development, with HSCs arising in the final wave in close association with the dorsal aorta. Recent reports have suggested that murine HSCs derive from haemogenic endothelial cells (ECs) lining the aortic floor1,2. Additional in vitro studies have similarly indicated that the haematopoietic progeny of ESCs arise through intermediates with endothelial potential3,4. Here we have used the unique strengths of the zebrafish embryo to image directly the generation of HSCs from the ventral wall of the dorsal aorta. Using combinations of fluorescent reporter transgenes, confocal time-lapse microscopy and flow cytometry, we have identified and isolated the stepwise intermediates as aortic haemogenic endothelium transitions to nascent HSCs. Finally, using a permanent lineage tracing strategy, we demonstrate that the HSCs generated from haemogenic endothelium are the lineal founders of the adult haematopoietic system.

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Figure 1: Direct imaging of HSC emergence from the embryonic aortic floor.
Figure 2: Prospective isolation of aortic haemogenic endothelium and nascent HSCs.
Figure 3: Long-term lineage tracing of embryonic endothelial cells.

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Acknowledgements

We thank S. Lin for providing kdrl:RFP animals. J.Y.B. was supported by the Irvington program of the Cancer Research Institute and by the California Institute for Regenerative Medicine (CIRM), N.C.C. by National Institutes of Health (NIH) HL074891, a Research and Education Foundation Award from GlaxoSmithKline and a Beginning Grant in Aid Award from the American Heart Association, B.S. by NIH F32DK752433, D.Y.R.S. by the Packard Foundation and NIH HL54737, and D.T. by a Scholar Award from the American Society of Hematology, a New Investigator Award from CIRM, and NIH DK074482.

Author Contributions J.Y.B., N.C.C. and D.T. designed experiments. J.Y.B. and D.T. wrote the manuscript, with key input from N.C.C. and D.Y.R.S.; J.Y.B. performed experiments. B.S. and S.T. generated and characterized the bactin:switch reporter line. N.C.C. and D.Y.R.S. generated kdrl:Cre and kdrl:memCherry transgenic lines.

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Author notes

  1. Julien Y. Bertrand and Neil C. Chi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cellular and Molecular Medicine,,

    Julien Y. Bertrand, Buyung Santoso, Shutian Teng & David Traver

  2. Section of Cell and Developmental Biology,,

    Julien Y. Bertrand, Buyung Santoso, Shutian Teng & David Traver

  3. Department of Medicine, University of California, San Diego, La Jolla, California 92093-0380, USA,

    Neil C. Chi

  4. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California 94158, USA,

    Neil C. Chi & Didier Y. R. Stainier

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  1. Julien Y. Bertrand
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Correspondence to David Traver.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-3 with Legends. (PDF 2510 kb)

Supplementary Movie 1

This movie shows emergence of HSCs from the floor of the dorsal aorta. (MOV 5321 kb)

Supplementary Movie 2

This movie shows close up of HSC emergence. (MOV 6540 kb)

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Bertrand, J., Chi, N., Santoso, B. et al. Haematopoietic stem cells derive directly from aortic endothelium during development. Nature 464, 108–111 (2010). https://doi.org/10.1038/nature08738

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