Embryonic Stem Cell–Derived CD166+ Precursors Develop Into Fully Functional Sinoatrial-Like Cells

• Published in: Circulation research Vol. 113; no. 4; pp. 389 - 398
• Main Authors: Scavone, Angela, Capilupo, Daniela, Mazzocchi, Nausicaa, Crespi, Alessia, Zoia, Stefano, Campostrini, Giulia, Bucchi, Annalisa, Milanesi, Raffaella, Baruscotti, Mirko, Benedetti, Sara, Antonini, Stefania, Messina, Graziella, DiFrancesco, Dario, Barbuti, Andrea
• Format: Journal Article
• Language: English
• Published: United States American Heart Association, Inc 02.08.2013
• Subjects: Acetylcholine - pharmacology; Activated-Leukocyte Cell Adhesion Molecule - metabolism; Animals; Biomarkers - metabolism; Cardiotonic Agents - pharmacology; Cell Differentiation - physiology; Cell Line; Cell Proliferation; Coculture Techniques; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Heart Ventricles - cytology; Heart Ventricles - drug effects; Heart Ventricles - metabolism; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels - metabolism; Isoproterenol - pharmacology; Mice; Models, Animal; Myocytes, Cardiac - cytology; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Sinoatrial Node - cytology; Sinoatrial Node - drug effects; Sinoatrial Node - metabolism; Stem Cells - cytology; Stem Cells - drug effects; Stem Cells - metabolism; sinoatrial node; cardiac progenitor cells; pacemaker; embryonic stem cells; HCN channels
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/CIRCRESAHA.113.301283

RATIONALE:A cell-based biological pacemaker is based on the differentiation of stem cells and the selection of a population displaying the molecular and functional properties of native sinoatrial node (SAN) cardiomyocytes. So far, such selection has been hampered by the lack of proper markers. CD166 is specifically but transiently expressed in the mouse heart tube and sinus venosus, the prospective SAN. OBJECTIVE:We have explored the possibility of using CD166 expression for isolating SAN progenitors from differentiating embryonic stem cells. METHODS AND RESULTS:We found that in embryonic day 10.5 mouse hearts, CD166 and HCN4, markers of the pacemaker tissue, are coexpressed. Sorting embryonic stem cells for CD166 expression at differentiation day 8 selects a population of pacemaker precursors. CD166 cells express high levels of genes involved in SAN development (Tbx18, Tbx3, Isl-1, Shox2) and function (Cx30.2, HCN4, HCN1, CaV1.3) and low levels of ventricular genes (Cx43, Kv4.2, HCN2, Nkx2.5). In culture, CD166 cells form an autorhythmic syncytium composed of cells morphologically similar to and with the electrophysiological properties of murine SAN myocytes. Isoproterenol increases (+57%) and acetylcholine decreases (−23%) the beating rate of CD166-selected cells, which express the β-adrenergic and muscarinic receptors. In cocultures, CD166-selected cells are able to pace neonatal ventricular myocytes at a rate faster than their own. Furthermore, CD166 cells have lost pluripotency genes and do not form teratomas in vivo. CONCLUSIONS:We demonstrated for the first time the isolation of a nonteratogenic population of cardiac precursors able to mature and form a fully functional SAN-like tissue.