Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Postnatal isl1+ cardioblasts enter fully differentiated cardiomyocyte lineages

Nature volume 433, pages 647–653 (2005)Cite this article

A Corrigendum to this article was published on 19 April 2007

Abstract

The purification, renewal and differentiation of native cardiac progenitors would form a mechanistic underpinning for unravelling steps for cardiac cell lineage formation, and their links to forms of congenital and adult cardiac diseases1,2,3. Until now there has been little evidence for native cardiac precursor cells in the postnatal heart4. Herein, we report the identification of isl1+ cardiac progenitors in postnatal rat, mouse and human myocardium. A cardiac mesenchymal feeder layer allows renewal of the isolated progenitor cells with maintenance of their capability to adopt a fully differentiated cardiomyocyte phenotype. Tamoxifen-inducible Cre/lox technology enables selective marking of this progenitor cell population including its progeny, at a defined time, and purification to relative homogeneity. Co-culture studies with neonatal myocytes indicate that isl1+ cells represent authentic, endogenous cardiac progenitors (cardioblasts) that display highly efficient conversion to a mature cardiac phenotype with stable expression of myocytic markers (25%) in the absence of cell fusion, intact Ca2+-cycling, and the generation of action potentials. The discovery of native cardioblasts represents a genetically based system to identify steps in cardiac cell lineage formation and maturation in development and disease.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

Buy

All prices are NET prices.

Figure 1: Isl1+ progenitors in the late embryonic and postnatal heart.
Figure 2: Genetic marking of isl1+ progenitors and myocytic cell fate.
Figure 3: Specificity of recombination and distribution of β-gal+ cells in isl1-mER-Cre-mER/R26R hearts.
Figure 4: Amplification, characterization and myocytic differentiation of isl1+ cardioblasts in vitro.
Figure 5: Real time [Ca2+]i transients and action potentials in FACS-sorted β-gal+ precursors after myocytic differentiation in co-culture.

References

  1. Chien, K. R. Stem cells: lost in translation. Nature 428, 607–608 (2004)

    Article  ADS  CAS  Google Scholar 

  2. Epstein, J. A. Developing models of DiGeorge syndrome. Trends Genet. 17, S13–S17 (2001)

    Article  CAS  Google Scholar 

  3. Chien, K. R. & Olson, E. C. Converging pathways and principles in heart development and disease: CV@CSH. Cell 110, 153–162 (2002)

    Article  CAS  Google Scholar 

  4. Rosenthal, N. Prometheus's vulture and the stem cell promise. N. Engl. J. Med. 346, 267–274 (2003)

    Article  Google Scholar 

  5. Bischoff, R. in Myogenesis (eds Engel, A. G. & Franszini-Armstrong, C.) 97–118 (McGraw-Hill, New York, 1994)

    Google Scholar 

  6. Seale, P. & Rudnicki, M. A. A new look at the origin, function, and “stem-cell” status of muscle satellite cells. Dev. Biol. 218, 115–124 (2000)

    Article  CAS  Google Scholar 

  7. Beltrami, A. P. et al. Adult cardiac stem cells are multipotent and support myocardial regeneration. Cell 114, 763–776 (2003)

    Article  CAS  Google Scholar 

  8. Oh, H. et al. Cardiac progenitor cells from adult myocardium: Homing, differentiation, and fusion after infarction. Proc. Natl Acad. Sci. USA 100, 12313–12318 (2003)

    Article  ADS  CAS  Google Scholar 

  9. Balsam, L. B. et al. Haematopoietic stem cells adopt mature haematopoietic fates in ischemic myocardium. Nature 428, 668–675 (2004)

    Article  ADS  CAS  Google Scholar 

  10. Murry, C. E. et al. Hematopoietic stem cells do not transdifferentiate into cardiac myocytes in myocardial infarcts. Nature 428, 664–668 (2004)

    Article  ADS  CAS  Google Scholar 

  11. Nygren, J. M. et al. Bone marrow-derived hematopoietic cells generate cardiomyocytes at a low-frequency through cell fusion, but not transdifferentiation. Nature Med. 10, 494–501 (2004)

    Article  CAS  Google Scholar 

  12. Cai, C.-L. et al. Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev. Cell 5, 877–889 (2003)

    Article  CAS  Google Scholar 

  13. Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001)

    Article  CAS  Google Scholar 

  14. Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)

    Article  CAS  Google Scholar 

  15. Verrou, C., Zhang, Y., Zurn, C., Schamel, W. W. & Reth, M. Comparison of the tamoxifen regulated chimeric Cre recombinases MerCreMer and CreMer. Biol. Chem. 380, 1435–1438 (1999)

    Article  CAS  Google Scholar 

  16. Danielian, P. S., Muccino, D., Rowitch, D. H., Michael, S. K. & McMahon, A. P. Modification of gene activity in mouse embryos in utero by tamoxifen-inducible form of Cre recombinase. Curr. Biol. 8, 1323–1326 (1998)

    Article  CAS  Google Scholar 

  17. Schwenk, F., Kuhn, R., Angrand, P. O., Rajewsky, K. & Stewart, A. F. Temporally and spatially regulated somatic mutagenesis in mice. Nucleic Acids Res. 26, 1427–1432 (1998)

    Article  CAS  Google Scholar 

  18. Martin, C. M. et al. Persistent expression of the ATP-binding cassette transporter, Abcg2, identifies cardiac SP cells in the developing and adult heart. Dev. Biol. 265, 262–275 (2004)

    Article  CAS  Google Scholar 

  19. Wurmser, A. E. et al. Cell fusion-independent differentiation of neural stem cells to the endothelial lineage. Nature 430, 350–356 (2004)

    Article  ADS  CAS  Google Scholar 

  20. Voura, E. B., Jaiswal, J. K., Mattoussi, H. & Simon, S. M. Tracking metastatic tumor cell extravasation with quantum dot nanocrystals and fluorescence emission-scanning microscopy. Nature Med. 9, 993–998 (2004)

    Article  Google Scholar 

  21. Maltsev, V. A., Ji, G. J., Wobus, A. M., Fleischmann, B. K. & Hescheler, J. Establishment of β-adrenergic modulation of L-type Ca2+ current in the early stages of cardiomyocyte development. Circ. Res. 84, 136–145 (1999)

    Article  CAS  Google Scholar 

  22. He, J.-Q., Ma, Y., Lee, Y., Thomson, J. A. & Kamp, T. J. Human embryonic stem cells develop into multiple types of cardiac myocytes. Circ. Res. 93, 32–39 (2003)

    Article  CAS  Google Scholar 

  23. D'Amour, K. A. & Gage, F. H. Are somatic stem cells pluripotent or lineage-restricted? Nature Med. 8, 213–214 (2002)

    Article  CAS  Google Scholar 

  24. Pashmforoush, M. et al. Nkx2.5 pathways and congenital heart disease: loss of ventricular myocyte lineage specification leads to progressive cardiomyopathy and complete heart block. Cell 117, 373–386 (2004)

    Article  CAS  Google Scholar 

  25. Hayashi, S. & McMahon, A. P. Efficient recombination in diverse tissues by tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. Dev. Biol. 244, 305–318 (2002)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank W. Giles for discussions and comments on the real-time Ca2+ measurements and electrophysiology, the National Center for Microscopy and Imaging Research at UCSD for the use of the high-speed multi-photon laser-scanning microscope (Bio-Rad RTS2000) and custom plug-ins for the ImageJ software (H. Hakozaki, S. Chow and M. Ellisman), the UCSD Cancer Center Digital Imaging Shared Resource for deconvolution microscopy (S. McMullen and J. Feramisco). We would like especially to acknowledge D. Young for his expertise and help in the FACS sorting analysis (UCSD Cancer Center Flow Cytometry Laboratories). This work was supported by the NIH and the Jean Le Ducq Foundation. K.-L.L. is a Heisenberg Scholar of the German Research Foundation.

Author information

Author notes

  1. Karl-Ludwig Laugwitz, Alessandra Moretti and Jason Lam: These authors contributed equally to this work

Authors and Affiliations

  1. Institute of Molecular Medicine,

    Karl-Ludwig Laugwitz, Alessandra Moretti, Jason Lam, Yinhong Chen, Sarah Woodard, Li-Zhu Lin, Chen-Leng Cai, Sylvia Evans & Kenneth R. Chien

  2. Department of Medicine, University of California, San Diego, School of Medicine, La Jolla, California, 92093, USA

    Oleksandr Platoshyn & Jason X.-J. Yuan

  3. Children's Hospital of Philadelphia, Cardiac Center, Philadelphia, Pennsylvania, 19104, USA

    Peter Gruber

  4. Cardiovascular Division, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA

    Min Min Lu

  5. Max-Planck Institut für Immunbiologie, Universität Freiburg, Biologie III, Abteilung Molekulare Immunologie, 79108, Freiburg, Germany

    Michael Reth

Authors
  1. Karl-Ludwig Laugwitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alessandra Moretti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jason Lam
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter Gruber
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yinhong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sarah Woodard
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Li-Zhu Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chen-Leng Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Min Min Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Michael Reth
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Oleksandr Platoshyn
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jason X.-J. Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Sylvia Evans
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Kenneth R. Chien
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Sylvia Evans or Kenneth R. Chien.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Table 1

Evaluation of human, post-natal isl1+ progenitors in atrial tissue following pediatric surgical correction of diverse congenital heart defects. (DOC 33 kb)

Supplementary Table 2

Comparison of isl1+ cardioblasts, cardiac sca-1+ cells and cardiac side population (SP) cells. (DOC 39 kb)

Supplementary Figure 1

Rare subset of isl1+ cardiac progenitors in double heterozygous isl1-mER-Cre-mER/R26R hearts in 2 month old animals in the outflow tract (a) and the right atrium (b). (PDF 133 kb)

Supplementary Figure 2

Amplification of rat isl1+ progenitors on cardiac mesenchymal cells in vitro (a, b) and their phenotypic characterization (c). (PDF 116 kb)

Supplementary Video

Video demonstrating the [Ca2+]i transients of differentiated β-Gal+ progenitors. (MOV 9207 kb)

Supplementary Video legend

Real time calcium imaging of FACS-sorted β-Gal+ progenitors differentiated in coculture. (DOC 21 kb)

Supplementary Method Section

Methods regarding flow cytometry analysis, co-culture experiments, immunocytochemistry and lacZ staining, RT-PCR, detection of [Ca2+]i transients and electrophysiological action potential recordings are described in the supplementary method section. (DOC 35 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Laugwitz, KL., Moretti, A., Lam, J. et al. Postnatal isl1+ cardioblasts enter fully differentiated cardiomyocyte lineages. Nature 433, 647–653 (2005). https://doi.org/10.1038/nature03215

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature03215

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing