BioProject

Analysis of gene expression of mESC-derived cardiac purkinje fiber-like cells at mRNA level. More...

Analysis of gene expression of mESC-derived cardiac purkinje fiber-like cells at mRNA level. Dysfunction of the cardiac conduction system (CCS) significantly impacts pathogenesis of arrhythmia, a major cause of morbidity and mortality. Strategies to derive cardiac conduction cells including Purkinje fiber cells (PC) 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 PC from cardiac progenitors initially expressing cardiac myosin heavy chain, and that it does so by activating cAMP signaling. These findings provide a novel strategy to derive scalable PC, along with insight into the ontogeny of CCS development. Overall design: Total RNA isolated from mESC-derived cardiac purkinje fiber-like cells. Mouse Contactin2:eGFP transduced with Lenti-viral aMHC: mCherry embryonic Stem Cells was used. Day 4 of differentiated cells were treated with Sodium Nitroprusside(SN). After 25 days differentiation, by using FACS, we could separate SN-induced cells into three populations: negative, αMhc:mCherry+ (MHC) and Cntn2:eGFP+/dim αMhc:mCherry+(GFP). We then sequenced mRNA from these three populations and found that Cntn2:eGFP+ cells express cardiac purkinje-fiber gene profiles. Less...