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A HCN4+ cardiomyogenic progenitor derived from the first heart field and human pluripotent stem cells

Nature Cell Biology volume 15, pages 10981106 (2013) | Download Citation


Most of the mammalian heart is formed from mesodermal progenitors in the first and second heart fields (FHF and SHF), whereby the FHF gives rise to the left ventricle and parts of the atria and the SHF to the right ventricle, outflow tract and parts of the atria1,2,3. Whereas SHF progenitors have been characterized in detail, using specific molecular markers4,5,6,7,8, comprehensive studies on the FHF have been hampered by the lack of exclusive markers. Here, we present Hcn4 (hyperpolarization-activated cyclic nucleotide-gated channel 4) as an FHF marker. Lineage-traced Hcn4+/FHF cells delineate FHF-derived structures in the heart and primarily contribute to cardiomyogenic cell lineages, thereby identifying an early cardiomyogenic progenitor pool. As a surface marker, HCN4 also allowed the isolation of cardiomyogenic Hcn4+/FHF progenitors from human embryonic stem cells. We conclude that a primary purpose of the FHF is to generate cardiac muscle and support the contractile activity of the primitive heart tube, whereas SHF-derived progenitors contribute to heart cell lineage diversification.

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We would like to thank C. Cowan for his support, L. Prickett-Rice, K. Folz-Donahue and M. Weglarz of the Harvard Stem Cell Institute Flow Cytometry Core Facility for assistance with FACS analysis, C. Du of the Tufts Electrophysiology Core for assistance with electrophysiology recordings, L. Bu for technical advice, K. Buac, E. Hansson, C. Riedel and L. Bu for critical reading of the manuscript and discussions, and C. Hartmann for advice on double-fluorescence in situ hybridizations. D.S. has received a D.F.G. (German Research Foundation) postdoctoral fellowship. K.B. was supported by a NHLBI T32HL007208 grant. This work is financially supported by the NIH U01 HL098 166 and NIH U01H100408 research grants.

Author information

Author notes

    • Monika K. Abramczuk
    • , Kristina Buac
    •  & Lior Zangi

    Present addresses: Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr Bohr-Gasse 3, 1030 Vienna, Austria (M.K.A.); Department of Genetics, University of Georgia, Athens, Georgia 30602, USA (K.B.); Department of Cardiology, Children’s Hospital Boston, Boston 02115, USA, and Harvard Stem Cell Institute, Cambridge, Massachusetts, USA (L.Z.)


  1. Cardiovascular Research Center, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA

    • Daniela Später
    • , Monika K. Abramczuk
    • , Kristina Buac
    • , Lior Zangi
    • , Maxine W. Stachel
    • , Jonathan Clarke
    • , Makoto Sahara
    •  & Kenneth R. Chien
  2. Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Medical School, 7 Divinity Avenue, Cambridge, Massachusetts 02138, USA

    • Daniela Später
    • , Monika K. Abramczuk
    • , Kristina Buac
    • , Lior Zangi
    • , Maxine W. Stachel
    • , Makoto Sahara
    •  & Kenneth R. Chien
  3. Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Friedrich-Alexander-Universität, Erlangen-Nürnberg, 91054 Erlangen, Germany

    • Andreas Ludwig
  4. Department of Cell and Molecular Biology and Medicine, Karolinska Institutet, 7 Berzelius Vag, Stockholm, Sweden

    • Kenneth R. Chien


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D.S. and K.R.C. designed the experiments and wrote the manuscript. D.S. carried out most of the experiments and analysed most of the data. M.K.A. helped to perform and analyse clonal analysis experiments. K.B. contributed PCRs with reverse transcription of in vivo clonal analysis. L.Z. performed some immunohistochemical staining, M.W.S. helped with some hESC differentiation assays and RNA isolations, J.C. performed acetylcholinesterase staining, M.S. helped with some mESC clonal analysis cultures and A.L. provided the Hcn4CreErt2 mouse line.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Kenneth R. Chien.

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  1. 1.

    Clonal analysis of single Hcn4+/FHF cells isolated from mouse embryos.

    Representative example of a beating cardiomyogenic clone derived from a single Hcn4+/FHF cell from mouse embryos.

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