Efficient Generation of Cardiac Purkinje Cells from ESCs by Activating cAMP Signaling

•A chemical screen was carried out for compounds that induce cardiac conduction cells•Two ESC reporter lines were used to identify lead hits•Sodium nitroprusside efficiently generated scalable amounts of PC-like cells•By activating cAMP signaling, PCs are derived from cardiac progenitors

SummaryDysfunction of the specialized cardiac conduction system (CCS) is associated with life-threatening arrhythmias. Strategies to derive CCS cells, including rare Purkinje cells (PCs), would facilitate models for mechanistic studies and drug discovery and also provide new cellular materials for regenerative therapies. A high-throughput chemical screen using CCS:lacz and Contactin2:egfp (Cntn2:egfp) reporter embryonic stem cell (ESC) lines was used to discover a small molecule, sodium nitroprusside (SN), that efficiently promotes the generation of cardiac cells that express gene profiles and generate action potentials of PC-like cells. Imaging and mechanistic studies suggest that SN promotes the generation of PCs from cardiac progenitors initially expressing cardiac myosin heavy chain and that it does so by activating cyclic AMP signaling. These findings provide a strategy to derive scalable PCs, along with insight into the ontogeny of CCS development.