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Continuous single-cell imaging of blood generation from haemogenic endothelium


Despite decades of research, the identity of the cells generating the first haematopoietic cells in mammalian embryos is unknown1. Indeed, whether blood cells arise from mesodermal cells, mesenchymal progenitors, bipotent endothelial–haematopoietic precursors or haemogenic endothelial cells remains controversial2,3,4,5,6,7,8,9. Proximity of endothelial and blood cells at sites of embryonic haematopoiesis, as well as their similar gene expression, led to the hypothesis of the endothelium generating blood. However, owing to lacking technology10 it has been impossible to observe blood cell emergence continuously at the single-cell level, and the postulated existence of haemogenic endothelial cells remains disputed1. Here, using new imaging and cell-tracking methods, we show that embryonic endothelial cells can be haemogenic. By continuous long-term single-cell observation of mouse mesodermal cells generating endothelial cell and blood colonies, it was possible to detect haemogenic endothelial cells giving rise to blood cells. Living endothelial and haematopoietic cells were identified by simultaneous detection of morphology and multiple molecular and functional markers. Detachment of nascent blood cells from endothelium is not directly linked to asymmetric cell division, and haemogenic endothelial cells are specified from cells already expressing endothelial markers. These results improve our understanding of the developmental origin of mammalian blood and the potential generation of haematopoietic stem cells from embryonic stem cells.

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Figure 1: ESC-derived mesodermal cells give rise to haemogenic endothelial colonies.
Figure 2: Continuous observation of endothelial and haematopoietic molecular markers.
Figure 3: Nascent blood cells generated from ECs express CD41 before CD45.
Figure 4: Mesodermal cells derived from mouse embryos give rise to haemogenic endothelial colonies.


  1. Godin, I. & Cumano, A. The hare and the tortoise: an embryonic haematopoietic race. Nature Rev. Immunol. 2, 593–604 (2002)

    CAS  Article  Google Scholar 

  2. 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 

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

    CAS  Article  Google Scholar 

  4. Choi, K., Kennedy, M., Kazarov, A., Papadimitriou, J. C. & Keller, G. A common precursor for hematopoietic and endothelial cells. Development 125, 725–732 (1998)

    CAS  Article  Google Scholar 

  5. de Bruijn, M. F. et al. Hematopoietic stem cells localize to the endothelial cell layer in the midgestation mouse aorta. Immunity 16, 673–683 (2002)

    CAS  Article  Google Scholar 

  6. 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  Article  Google Scholar 

  7. Jaffredo, T. et al. From hemangioblast to hematopoietic stem cell: an endothelial connection? Exp. Hematol. 33, 1029–1040 (2005)

    Article  Google Scholar 

  8. Nishikawa, S. I. et al. In vitro generation of lymphohematopoietic cells from endothelial cells purified from murine embryos. Immunity 8, 761–769 (1998)

    CAS  Article  Google Scholar 

  9. Ueno, H. & Weissman, I. L. Clonal analysis of mouse development reveals a polyclonal origin for yolk sac blood islands. Dev. Cell 11, 519–533 (2006)

    CAS  Article  Google Scholar 

  10. Schroeder, T. Imaging stem cell driven mammalian regeneration. Nature 453, 345–351 (2008)

    ADS  CAS  Article  Google Scholar 

  11. Fraser, S. T. et al. Definitive hematopoietic commitment within the embryonic vascular endothelial-cadherin(+) population. Exp. Hematol. 30, 1070–1078 (2002)

    CAS  Article  Google Scholar 

  12. Sugiyama, D. et al. Erythropoiesis from acetyl LDL incorporating endothelial cells at the preliver stage. Blood 101, 4733–4738 (2003)

    CAS  Article  Google Scholar 

  13. Schroeder, T. et al. Recombination signal sequence-binding protein Jκ alters mesodermal cell fate decisions by suppressing cardiomyogenesis. Proc. Natl Acad. Sci. USA 100, 4018–4023 (2003)

    ADS  CAS  Article  Google Scholar 

  14. Schroeder, T. et al. Activated Notch1 alters differentiation of embryonic stem cells into mesodermal cell lineages at multiple stages of development. Mech. Dev. 123, 570–579 (2006)

    CAS  Article  Google Scholar 

  15. Hirashima, M., Kataoka, H., Nishikawa, S., Matsuyoshi, N. & Nishikawa, S. Maturation of embryonic stem cells into endothelial cells in an in vitro model of vasculogenesis. Blood 93, 1253–1263 (1999)

    CAS  Article  Google Scholar 

  16. Keller, G. Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev. 19, 1129–1155 (2005)

    CAS  Article  Google Scholar 

  17. Nakano, T., Kodama, H. & Honjo, T. In vitro development of primitive and definitive erythrocytes from different precursors. Science 272, 722–724 (1996)

    ADS  CAS  Article  Google Scholar 

  18. Huber, T. L., Kouskoff, V., Fehling, H. J., Palis, J. & Keller, G. Haemangioblast commitment is initiated in the primitive streak of the mouse embryo. Nature 432, 625–630 (2004)

    ADS  CAS  Article  Google Scholar 

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

    ADS  CAS  Article  Google Scholar 

  20. Kataoka, H. et al. Expressions of PDGF receptor alpha, c-Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells. Dev. Growth Differ. 39, 729–740 (1997)

    CAS  Article  Google Scholar 

  21. Shalaby, F. et al. A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. Cell 89, 981–990 (1997)

    CAS  Article  Google Scholar 

  22. Yamashita, J. et al. Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors. Nature 408, 92–96 (2000)

    ADS  CAS  Article  Google Scholar 

  23. Guo, R., Sakamoto, H., Sugiura, S. & Ogawa, M. Endothelial cell motility is compatible with junctional integrity. J. Cell. Physiol. 211, 327–335 (2007)

    CAS  Article  Google Scholar 

  24. Lampugnani, M. G. et al. A novel endothelial-specific membrane protein is a marker of cell–cell contacts. J. Cell Biol. 118, 1511–1522 (1992)

    CAS  Article  Google Scholar 

  25. Voyta, J. C., Via, D. P., Butterfield, C. E. & Zetter, B. R. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J. Cell Biol. 99, 2034–2040 (1984)

    CAS  Article  Google Scholar 

  26. Morita, K., Sasaki, H., Furuse, M. & Tsukita, S. Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells. J. Cell Biol. 147, 185–194 (1999)

    CAS  Article  Google Scholar 

  27. Taoudi, S. et al. Extensive hematopoietic stem cell generation in the AGM region via maturation of VE-cadherin+CD45+ pre-definitive HSCs. Cell Stem Cell 3, 99–108 (2008)

    CAS  Article  Google Scholar 

  28. Ferkowicz, M. J. et al. CD41 expression defines the onset of primitive and definitive hematopoiesis in the murine embryo. Development 130, 4393–4403 (2003)

    CAS  Article  Google Scholar 

  29. 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 

  30. Mitjavila-Garcia, M. T. et al. Expression of CD41 on hematopoietic progenitors derived from embryonic hematopoietic cells. Development 129, 2003–2013 (2002)

    CAS  Article  Google Scholar 

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We are grateful to S. Nishikawa, M. A. Rieger, A. Hermann and M. Yoder for technical advice and discussions, M. Ogawa and H. Niwa for the VEVC2 and EB3 ESC lines, respectively, B. Schauberger for programming contributions, and A. Roth and C. Raithel for technical support. We thank M. Goetz, A. Hermann, M. A. Rieger and A. IJpenberg for critical reading of the manuscript. Part of this study was financed by the Deutsche Forschungsgemeinschaft to T.S. and by the Leading Project for the Realization of Regenerative Medicine to S.-I.N.

Author Contributions H.M.E. planned and performed experiments; S.-I.N. discussed results and commented on the manuscript; T.S. designed the study and experiments, developed the time-lapse imaging and cell-tracking technology, performed initial experiments, analysed data with H.M.E. and wrote the paper with H.M.E.

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Correspondence to Timm Schroeder.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-8 with Legends, Supplementary Tables 1-3 and Supplementary Movies Legends 1-8 (PDF 2443 kb)

Supplementary Video 1

This time-lapse movie, covering a period of about 4.5 days, shows that cells with endothelial morphology can give rise to blood cells (see file s1 for full legend). (MOV 9773 kb)

Supplementary Video 2

This time-lapse movie, covering a period of about 1 day, proofs the cell identity of endothelial and blood cells by molecular and functional markers (see file s1 for full legend). (MOV 3788 kb)

Supplementary Video 3

This time-lapse movie, covering a period of about 2.5 days, shows that nascent blood cells generated by endothelial cells express the surface marker CD41 prior to CD45 expression (see file s1 for full legend). (MOV 4070 kb)

Supplementary Video 4

This time-lapse movie, covering a period of about 1.5 days, shows that mesodermal cells purified from 7.5 dpc mouse embryos can generate hemogenic endothelial cells (see file s1 for full legend). (MOV 3892 kb)

Supplementary Video 5

This time-lapse movie, covering a period of about 6.5 days, shows that nascent blood cells generated by endothelial cells have colony forming potential with high proliferative potential and multilineage (including megakaryocytic) potential. (MOV 8815 kb)

Supplementary Video 6

This time-lapse movie, covering a period of about 5 hours, demonstrates the temporal and optical resolution of the primary image sequences used for cell tracking (see file s1 for full legend). (MOV 2331 kb)

Supplementary Video 7

This time-lapse movie, covering a period of about 8.5 hours, demonstrates the temporal and optical resolution of the primary image sequences used for cell tracking (see file s1 for full legend). (MOV 6633 kb)

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Eilken, H., Nishikawa, SI. & Schroeder, T. Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature 457, 896–900 (2009).

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