c-kit expression identifies cardiovascular precursors in the neonatal heart

• DOI: 10.1073/pnas.0808920106
• Publication Date: 2009-02-05T02:39:22Z
• Authors: Yvonne N. Tallini; Kai Su Greene; Michael Craven; Alyson Spealman; Martin Breitbach; James Smith; Patricia J. Fisher; Michele Steffey; Michael Hesse; Robert M. Doran; Ashley Woods; Babu Singh; Andrew Yen; Bernd K. Fleischmann; Michael I. Kotlikoff
• Keywords: 0303 health sciences; Multipotent Stem Cells; Myocardium; Green Fluorescent Proteins; Cell Differentiation; Mice, Inbred Strains; Mice, Transgenic; Embryo, Mammalian; Cardiovascular System; Coronary Vessels; Cryosurgery; Mesoderm; Mice; Proto-Oncogene Proteins c-kit; 03 medical and health sciences; Animals, Newborn; Animals; Cell Lineage; Myocytes, Cardiac

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