c-kit expression identifies cardiovascular precursors in the neonatal heart

Yvonne N. Tallini, Su Greene Kai, Michael Craven, Alyson Spealman, Martin Breitbach, James Smith, Patricia J. Fisher, Michele A Steffey, Michael Hesse, Robert M. Doran, Ashley Woods, Babu Singh, Andrew Yen, Bernd K. Fleischmann, Michael I. Kotlikoff

Research output: Contribution to journalArticle

155 Scopus citations

Abstract

Directed differentiation of embryonic stem cells indicates that mesodermal lineages in the mammalian heart (cardiac, endothelial, and smooth muscle cells) develop from a common, multipotent cardiovascular precursor. To isolate and characterize the lineage potential of a resident pool of cardiovascular progenitor cells (CPcs), we developed BAC transgenic mice in which enhanced green fluorescent protein (EGFP) is placed under control of the c-kit locus (c-kitBAC-EGFP mice). Discrete c-kit-EGFP+ cells were observed at different stages of differentiation in embryonic hearts, increasing in number to a maximum at about postnatal day (PN) 2; thereafter, EGFP + cells declined and were rarely observed in the adult heart. EGFP+ cells purified from PN 0-5 hearts were nestin+ and expanded in culture; 67% of cells were fluorescent after 9 days. Purified cells differentiated into endothelial, cardiac, and smooth muscle cells, and differentiation could be directed by specific growth factors. CPc-derived cardiac myocytes displayed rhythmic beating and action potentials characteristic of multiple cardiac cell types, similar to ES cell-derived cardiomyocytes. Single-cell dilution studies confirmed the potential of individual CPcs to form all 3 cardiovascular lineages. In adult hearts, cryoablation resulted in c-kit-EGFP+ expression, peaking 7 days postcryolesion. Expression occurred in endothelial and smooth muscle cells in the revascularizing infarct, and in terminally differentiated cardiomyocytes in the border zone surrounding the infarct. Thus, c-kit expression marks CPc in the neonatal heart that are capable of directed differentiation in vitro; however, c-kit expression in cardiomyocytes in the adult heart after injury does not identify cardiac myogenesis.

Original languageEnglish (US)
Pages (from-to)1808-1813
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number6
DOIs
StatePublished - Feb 10 2009
Externally publishedYes

Keywords

  • Bacterial artificial chromosome
  • Progenitor cell
  • Stem cell

ASJC Scopus subject areas

  • General

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    Tallini, Y. N., Kai, S. G., Craven, M., Spealman, A., Breitbach, M., Smith, J., Fisher, P. J., Steffey, M. A., Hesse, M., Doran, R. M., Woods, A., Singh, B., Yen, A., Fleischmann, B. K., & Kotlikoff, M. I. (2009). c-kit expression identifies cardiovascular precursors in the neonatal heart. Proceedings of the National Academy of Sciences of the United States of America, 106(6), 1808-1813. https://doi.org/10.1073/pnas.0808920106