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Human ISL1 heart progenitors generate diverse multipotent cardiovascular cell lineages

Abstract

The generation and expansion of diverse cardiovascular cell lineages is a critical step during human cardiogenesis, with major implications for congenital heart disease. Unravelling the mechanisms for the diversification of human heart cell lineages has been hampered by the lack of genetic tools to purify early cardiac progenitors and define their developmental potential1,2,3,4. Recent studies in the mouse embryo have identified a multipotent cardiac progenitor that contributes to all of the major cell types in the murine heart5,6,7,8. In contrast to murine development, human cardiogenesis has a much longer onset of heart cell lineage diversification and expansion, suggesting divergent pathways. Here we identify a diverse set of human fetal ISL1+ cardiovascular progenitors that give rise to the cardiomyocyte, smooth muscle and endothelial cell lineages. Using two independent transgenic and gene-targeting approaches in human embryonic stem cell lines, we show that purified ISL1+ primordial progenitors are capable of self-renewal and expansion before differentiation into the three major cell types in the heart. These results lay the foundation for the generation of human model systems for cardiovascular disease and novel approaches for human regenerative cardiovascular medicine.

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Figure 1: Analysis of the in vivo expression of ISL1 in SHF-derived structures of the human fetal heart.
Figure 2: ISL1 marks human ES-cell-derived cardiac progenitors.
Figure 3: Isolation and characterization of human ES-cell-derived ISL1 + cardiac progenitors and their progeny.
Figure 4: Expansion of human ES-cell-derived ISL1 + cardiac progenitors on Wnt3a feeders.

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Acknowledgements

We thank C. Cowan for advice on human ES cell culture and electroporation; A. Nagy for providing the DsRed-MST plasmid; Y. Qyang for discussion and comments; E. Hansson for a critical reading of this manuscript; M. Lindsay and M. Ortega-Molina for help on human fetal heart anatomy; L. B. Prickett-Rice and K. Folz-Donahue for flow cytometry support; and Advanced Bioscience Resources for providing the human fetal tissues. S.M.P. is funded by Foundation Alfonso Martin Escudero, Spain. This study is supported by Harvard Stem Cell Institute and the Leducq Foundation.

Author Contributions L.B. and X.J. carried out most of the human ES cell experiments and analysed the data; S.M.-P. performed the analysis on the human fetal hearts, her contribution is similar in significance to the contributions of L.B. and X.J.; L.B. and K.R.C. designed the study; L.B., X.J., S.M.-P. and K.R.C. wrote the manuscript; L.C. performed a part of the differentiation study on ISL1-β-geo BAC transgenic cells and ISL1+ expansion using wild-type human ES cells; S.Z. and Y.S. helped culture human ES cells and carried out some experiments; D.J.R. provided a part of the human fetal samples; P.L.H. provided advice on experiments; and I.J.D. provided pCAG-flox-DsRed plasmid and revised the manuscript.

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Correspondence to Kenneth R. Chien.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-5 with Legends and Supplementary Table 1. (PDF 3213 kb)

Supplementary Movie 1

This movie file shows spontaneous contraction in human ISL1-cre DsRed knock-in cells. (MOV 811 kb)

Supplementary Movie 2

This movie file shows fluorescent contracting DsRed+ clusters. (MOV 141 kb)

Supplementary Movie 3

This movie file shows contracting DsRed+ clusters in bright fields. (MOV 2179 kb)

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Bu, L., Jiang, X., Martin-Puig, S. et al. Human ISL1 heart progenitors generate diverse multipotent cardiovascular cell lineages. Nature 460, 113–117 (2009). https://doi.org/10.1038/nature08191

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