Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population (original) (raw)

Nature volume 453, pages 524–528 (2008)Cite this article

Abstract

The functional heart is comprised of distinct mesoderm-derived lineages including cardiomyocytes, endothelial cells and vascular smooth muscle cells. Studies in the mouse embryo and the mouse embryonic stem cell differentiation model have provided evidence indicating that these three lineages develop from a common Flk-1+ (kinase insert domain protein receptor, also known as Kdr) cardiovascular progenitor that represents one of the earliest stages in mesoderm specification to the cardiovascular lineages1. To determine whether a comparable progenitor is present during human cardiogenesis, we analysed the development of the cardiovascular lineages in human embryonic stem cell differentiation cultures. Here we show that after induction with combinations of activin A, bone morphogenetic protein 4 (BMP4), basic fibroblast growth factor (bFGF, also known as FGF2), vascular endothelial growth factor (VEGF, also known as VEGFA) and dickkopf homolog 1 (DKK1) in serum-free media, human embryonic-stem-cell-derived embryoid bodies generate a KDRlow/C-KIT(CD117)neg population that displays cardiac, endothelial and vascular smooth muscle potential in vitro and, after transplantation, in vivo. When plated in monolayer cultures, these KDRlow/C-KITneg cells differentiate to generate populations consisting of greater than 50% contracting cardiomyocytes. Populations derived from the KDRlow/C-KITneg fraction give rise to colonies that contain all three lineages when plated in methylcellulose cultures. Results from limiting dilution studies and cell-mixing experiments support the interpretation that these colonies are clones, indicating that they develop from a cardiovascular colony-forming cell. Together, these findings identify a human cardiovascular progenitor that defines one of the earliest stages of human cardiac development.

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Acknowledgements

We thank M. Oza for MEA (multi-electrode arrays) measurement and members of the Keller laboratory for critically reading this manuscript. G.M.K., S.J.K. and G.W.A. are supported by the National Institutes of Health/National Heart Lung and Blood Institute.

Author Contributions L.Y. carried out most of the experiments; L.Y., S.J.K. and G.M.K. designed the study; L.Y. and G.M.K. analysed the data and wrote the manuscript; M.H.S. and L.J.F. performed the transplantation and differentiation study in the normal hearts; E.D.A. was responsible for the transplantation and analyses of the infracted hearts; T.K.R. and G.W.A. carried out the patch-clamp study; E.H. and R.M.L. generated the AAV (adeno-associated virus)–GFP–hES2 cells; M.K. provided advice on experimental design and analysed data; and L.Y. and K.B. performed the field potential recording.

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Authors and Affiliations

  1. Department of Gene and Cell Medicine, The Black Family Stem Cell Institute, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, New York 10029, USA,
    Lei Yang, Eric D. Adler, R. Michael Linden & Gordon M. Keller
  2. Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 West Walnut Street, Indiana 46202, USA ,
    Mark H. Soonpaa & Loren J. Field
  3. Greenberg Division of Cardiology, Departments of Medicine and Pharmacology, Weill Medical College of Cornell University, 520 East 70th Street, New York, New York 10021, USA,
    Torsten K. Roepke & Geoffrey W. Abbott
  4. McEwen Centre for Regenerative Medicine, University Health Network, 101 College Street, Toronto, Ontario M5G 1L7, Canada ,
    Steven J. Kattman, Marion Kennedy & Gordon M. Keller
  5. Department of Infectious Diseases, King’s College London, London SE1 9RT, UK
    Els Henckaerts & R. Michael Linden
  6. VistaGen Therapeutics Inc., 384 Oyster Point Boulevard, Suite 8, San Francisco, California 94080, USA ,
    Kristina Bonham

Authors

  1. Lei Yang
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  2. Mark H. Soonpaa
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  3. Eric D. Adler
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  4. Torsten K. Roepke
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  5. Steven J. Kattman
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  6. Marion Kennedy
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  7. Els Henckaerts
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  8. Kristina Bonham
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  9. Geoffrey W. Abbott
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  10. R. Michael Linden
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  11. Loren J. Field
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  12. Gordon M. Keller
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Corresponding author

Correspondence toGordon M. Keller.

Supplementary information

Supplementary information

The file contains Supplementary Methods, Supplementary Figures 1-5 with Legends and Legends to Supplementary Movies S1-S4. (PDF 4882 kb)

Supplementary information

The file contains Supplementary Movie S1 showing day 14 EBs with contracting cells derived from hES2 cells. (MOV 317 kb)

Supplementary information

The file contains Supplementary Movie S2 showing aggregates with contracting cells generated from day 6 EB-derived KDRlow/C-KITneg cells. Aggregates were cultured in the low cluster plates for 10 days (MOV 592 kb)

Supplementary information

The file contains Supplementary Movie S3 showing monolayer of contracting cells generated from day 6 EB-derived KDRlow/C-KITneg cells. The sorted cells were cultured on a gelatin coated well for 10 days (MOV 753 kb)

Supplementary information

The file contains Supplementary Movie S4 showing a cardiac colony generated from day 6 EB-derived KDRlow/C-KITneg cells. The colony was maintained in methylcellulose cultures for 10 days. (MOV 336 kb)

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Yang, L., Soonpaa, M., Adler, E. et al. Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population.Nature 453, 524–528 (2008). https://doi.org/10.1038/nature06894

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Editorial Summary

Embryonic stem cells: Take heart from a growth factor cocktail

A method for differentiation and isolation of one of the earliest human cardiac progenitors from human embryonic stem cells has been developed. By supplying a cocktail of growth factors at the appropriate stage of development, these cells can form cardiac, endothelial and vascular smooth muscle in vitro and, when transplanted, in vivo. When plated in culture, they can form populations of contracting cardiomyocytes. Transplantation of the cells into damaged mice hearts improved cardiac function. These cells will be useful for the study of cardiac development, and provide an enriched source of progenitors for engineering cardiovascular tissue in vitro and for transplantation to large animal models of heart disease.