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Haematopoietic stem cells derive directly from aortic endothelium during development

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.

References

  1. Zovein, A. C. et al. Fate tracing reveals the endothelial origin of hematopoietic stem cells. Cell Stem Cell 3, 625–636 (2008)

    CAS  Article  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)

    ADS  CAS  Article  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)

    ADS  CAS  Article  Google Scholar 

  4. Lancrin, C. et al. The haemangioblast generates haematopoietic cells through a haemogenic endothelium stage. Nature 457, 892–895 (2009)

    ADS  CAS  Article  Google Scholar 

  5. Cumano, A. & Godin, I. Ontogeny of the hematopoietic system. Annu. Rev. Immunol. 25, 745–785 (2007)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  Google Scholar 

  10. North, T. E. et al. Prostaglandin E2 regulates vertebrate haematopoietic stem cell homeostasis. Nature 447, 1007–1011 (2007)

    ADS  CAS  Article  Google Scholar 

  11. Chi, N. C. et al. Foxn4 directly regulates tbx2b expression and atrioventricular canal formation. Genes Dev. 22, 734–739 (2008)

    CAS  Article  Google Scholar 

  12. Kissa, K. et al. Live imaging of emerging hematopoietic stem cells and early thymus colonization. Blood 111, 1147–1156 (2008)

    CAS  Article  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)

    CAS  Article  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)

    ADS  CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  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)

    CAS  Article  Google Scholar 

  26. MacPherson, D. et al. Cell type-specific effects of Rb deletion in the murine retina. Genes Dev. 18, 1681–1694 (2004)

    CAS  Article  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)

    CAS  Article  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.

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Correspondence to David Traver.

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The authors declare no competing financial interests.

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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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