Haematopoietic stem cells derive directly from aortic endothelium during development (original) (raw)

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

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Zovein, A. C. et al. Fate tracing reveals the endothelial origin of hematopoietic stem cells. Cell Stem Cell 3, 625–636 (2008)
    Article CAS Google Scholar
  2. Chen, M. J., Yokomizo, T., Zeigler, B. M., Dzierzak, E. & Speck, N. A. Runx1 is required for the endothelial to haematopoietic cell transition but not thereafter. Nature 457, 887–891 (2009)
    Article ADS CAS Google Scholar
  3. Eilken, H. M., Nishikawa, S. & Schroeder, T. Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature 457, 896–900 (2009)
    Article ADS CAS Google Scholar
  4. Lancrin, C. et al. The haemangioblast generates haematopoietic cells through a haemogenic endothelium stage. Nature 457, 892–895 (2009)
    Article ADS CAS Google Scholar
  5. Cumano, A. & Godin, I. Ontogeny of the hematopoietic system. Annu. Rev. Immunol. 25, 745–785 (2007)
    Article CAS Google Scholar
  6. Murry, C. E. & Keller, G. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell 132, 661–680 (2008)
    Article CAS Google Scholar
  7. de Bruijn, M. F., Speck, N. A., Peeters, M. C. & Dzierzak, E. Definitive hematopoietic stem cells first develop within the major arterial regions of the mouse embryo. EMBO J. 19, 2465–2474 (2000)
    Article CAS Google Scholar
  8. Dzierzak, E. & Speck, N. A. Of lineage and legacy: the development of mammalian hematopoietic stem cells. Nature Immunol. 9, 129–136 (2008)
    Article CAS Google Scholar
  9. Bertrand, J. Y., Kim, A. D., Teng, S. & Traver, D. CD41+ cmyb+ precursors colonize the zebrafish pronephros by a novel migration route to initiate adult hematopoiesis. Development 135, 1853–1862 (2008)
    Article CAS Google Scholar
  10. North, T. E. et al. Prostaglandin E2 regulates vertebrate haematopoietic stem cell homeostasis. Nature 447, 1007–1011 (2007)
    Article ADS CAS Google Scholar
  11. Chi, N. C. et al. Foxn4 directly regulates tbx2b expression and atrioventricular canal formation. Genes Dev. 22, 734–739 (2008)
    Article CAS Google Scholar
  12. Kissa, K. et al. Live imaging of emerging hematopoietic stem cells and early thymus colonization. Blood 111, 1147–1156 (2008)
    Article CAS Google Scholar
  13. Huang, H., Zhang, B., Hartenstein, P. A., Chen, J. N. & Lin, S. NXT2 is required for embryonic heart development in zebrafish. BMC Dev. Biol. 5, 7 (2005)
    Article Google Scholar
  14. Mikkola, H. K., Fujiwara, Y., Schlaeger, T. M., Traver, D. & Orkin, S. H. Expression of CD41 marks the initiation of definitive hematopoiesis in the mouse embryo. Blood 101, 508–516 (2003)
    Article CAS Google Scholar
  15. Bertrand, J. Y. et al. Characterization of purified intraembryonic hematopoietic stem cells as a tool to define their site of origin. Proc. Natl Acad. Sci. USA 102, 134–139 (2005)
    Article ADS CAS Google Scholar
  16. Jin, S. W., Beis, D., Mitchell, T., Chen, J. N. & Stainier, D. Y. Cellular and molecular analyses of vascular tube and lumen formation in zebrafish. Development 132, 5199–5209 (2005)
    Article CAS Google Scholar
  17. Bussmann, J., Bakkers, J. & Schulte-Merker, S. Early endocardial morphogenesis requires Scl/Tal1. PLoS Genet. 3, e140 (2007)
    Article Google Scholar
  18. Liao, W. et al. The zebrafish gene cloche acts upstream of a flk-1 homologue to regulate endothelial cell differentiation. Development 124, 381–389 (1997)
    CAS PubMed Google Scholar
  19. Choi, J. et al. FoxH1 negatively modulates flk1 gene expression and vascular formation in zebrafish. Dev. Biol. 304, 735–744 (2007)
    Article CAS Google Scholar
  20. Traver, D. et al. Transplantation and in vivo imaging of multilineage engraftment in zebrafish bloodless mutants. Nature Immunol. 4, 1238–1246 (2003)
    Article CAS Google Scholar
  21. Jaffredo, T., Gautier, R., Eichmann, A. & Dieterlen-Lievre, F. Intraaortic hemopoietic cells are derived from endothelial cells during ontogeny. Development 125, 4575–4583 (1998)
    CAS PubMed Google Scholar
  22. Ciau-Uitz, A., Walmsley, M. & Patient, R. Distinct origins of adult and embryonic blood in Xenopus . Cell 102, 787–796 (2000)
    Article CAS Google Scholar
  23. Feng, H. et al. Heat-shock induction of T-cell lymphoma/leukaemia in conditional Cre/lox-regulated transgenic zebrafish. Br. J. Haematol. 138, 169–175 (2007)
    Article CAS Google Scholar
  24. Westerfield, M. The zebrafish book: A guide for the laboratory use of zebrafish (Brachydanio rerio) 2.1 edn (Univ. of Oregon Press, 1994)
    Google Scholar
  25. Beis, D. et al. Genetic and cellular analyses of zebrafish atrioventricular cushion and valve development. Development 132, 4193–4204 (2005)
    Article CAS Google Scholar
  26. MacPherson, D. et al. Cell type-specific effects of Rb deletion in the murine retina. Genes Dev. 18, 1681–1694 (2004)
    Article CAS Google Scholar
  27. Kawakami, K. et al. A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish. Dev. Cell 7, 133–144 (2004)
    Article CAS Google Scholar
  28. Rozen, S. & Skaletsky, H. J. Primer3 on the WWW for general users and biologist programmers. Methods Mol. Biol. 132, 365–386 (2000)
    CAS Google Scholar

Download references

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.

Author information

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

Authors

  1. Julien Y. Bertrand
    You can also search for this author inPubMed Google Scholar
  2. Neil C. Chi
    You can also search for this author inPubMed Google Scholar
  3. Buyung Santoso
    You can also search for this author inPubMed Google Scholar
  4. Shutian Teng
    You can also search for this author inPubMed Google Scholar
  5. Didier Y. R. Stainier
    You can also search for this author inPubMed Google Scholar
  6. David Traver
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toDavid Traver.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

PowerPoint slides

Rights and permissions

About this article

Cite this article

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

Download citation

This article is cited by

Editorial Summary

Blood stem cell creation

In zebrafish, haematopoietic stem cells (HSCs) arise from the dorsal aorta of the embryo. In vitro studies have suggested that there are in the dorsal aorta a population of intermediate progenitors that can give rise to both endothelial (or blood vessel lineage) and blood cells. In this issue, two groups present images showing the birth of HSCs from the ventral wall of the dorsal aorta in live zebrafish embryos. Bertrand et al. combined fluorescent reporter transgenes, confocal time-lapse microscopy and flow cytometry to identify and isolate the stepwise intermediates as aortic haemogenic endothelium transitions to nascent HSCs. They also show that the HSCs generated from this haemogenic endothelium are the lineal founders of virtually all of the adult haematopoietic system. Karima Kissa and Philippe Herbomel similarly use imaging of live zebrafish to show HSCs emerge directly from the aorta floor, They show this process that does not involve cell division but movement of single endothelial cells out of the aorta ventral wall into the sub-aortic space, where they transform into haematopoietic cells. They call this new type of cell behaviour endothelial haematopoietic transition (EHT). In a third report, Boisset et al. confirm that this process also occurs in mice, using a dissection procedure to visualize the deeply located aorta. They showed de novo emergence of phenotypically defined HSCs directly from ventral aortic haemogenic endothelial cells.