A novel cardiomyogenic role for Isl1+ neural crest cells in the inflow tract

The goal of this study was to analyze the cellular compsitions of the arterial pole and venous pole regions of the neonatal mouse heart using single-cell RNA sequencing. Overall design: The dorsal and ventral domains of the cardiac base were dissected from 3-day old C57Bl6 male and female neonatal mice (n=15) and enzymatically dissociated into single-cell suspensions. The ventral domain included the proximal aorta, pulmonary artery, and outflow tract base. The dorsal domain included the sinoatrial node, cardiac ganglia and pulmonary veins. Single-cell preparations from 15 neonatal hearts were combined to generate two pools of dorsal and ventral cells, and cultured in vitro for 5 days to allow recovery from enzymatic and mechanical stress. Subsequently, cells were processed for 3' single cell RNA-seq using the 10X Genomics Chromium platform and sequenced on Illumina Novaseq 6000.