Yifei Miao#, Lei Tian#, Marcy Martin, Sharon L. Paige, Francisco X. Galdos, Jibiao Li, Alyssa Klein, Hao Zhang, Ning Ma, Yuning Wei, Maria Stewart, Soah Lee, Jan-Renier Moonen, Bing Zhang, Paul Grossfeld, Seema Mital, David Chitayat, Joseph C. Wu, Marlene Rabinovitch, Timothy J. Nelson, Shuyi Nie, Sean M. Wu, Mingxia Gu. Single-Cell Transcriptomic Analysis Reveals Developmentally Impaired Endocardial Population in Hypoplastic Left Heart Syndrome.

#These authors contributed equally

Contact person: Mingxia Gu, Mingxia.Gu@cchmc.org

To unveil the complexity of human cardiac cell heterogeneity and comprehensive cell-cell interactions during the heart development, we applied single-cell RNA-seq to profile the transcriptome of a human fetal heart at gestational age day 83. We manually dissected the right-ventricular (RV) and left-ventricular (LV) free walls and enzymatically isolated and purified the endothelial cell (ECs) using CD144 microbeads-based MACS sorting. All cells (both CD144+ and CD144-) were then applied to 10X chromium 3’ single-cell RNA-seq platform (V2 version). Chromium Single Cell Software Suite v2.1.1 was used for pre-processing the single cell RNA-seq data. A substantial number of cells were captured in each sample: LV_EC (2633 cells), RV_EC (1890 cells), LV_nonEC (2739 cells), RV_nonEC (2738 cells). By cell clustering and annotation, major cardiac cell types were retrieved, including cardiomyocyte, endocardium, venous EC, arterial EC, capillary EC, lymphatic EC, pericyte, fibroblast, epicardium, immune cells, conduction system, and nervous system.