Multi-lineage cardiac model recapitulates human cardiogenesis and heart defects

Congenital heart defects constitute the leading cause of human fetal death, however the underlying mechanisms of most cardiac defects remain unknown. The ability to generate in vitro cardiac models mimicking significant aspects of native heart development is a prerequisite for disease modeling. Here, we established a 2D cardiac platform that equivalent of human fetal heart with pluripotent stem cells (hPSCs). Through the precise manipulation of retinoic acid (RA) signaling, we recapitulated the anterior-posterior patterning found in embryogenesis. By lineage tracing analyses and single-cell transcriptomic profiling, we described the lineage relationships between distinct populations and the developmental potentials of heart-field-specific progenitors. Comparisons of hPSC-derived and gastrulating human hearts enabled the identification of in vivo-like cardiac molecular characteristics in our platform. We used this model to unravel maternal pregestational diabetes-induced congenital heart defects. Together, the 2D cardiac platform offers a unique tool for disease modeling, drug discovery and cardiogenesis studying. Overall design: To investigate the transcriptomic profiles of populations generated under two distinct conditions, scRNA-seq was utilized during the in vitro differentiation of human pluripotent stem cells (hPSCs) into cardiac lineages. This process encompassed six key developmental stages (days 0, 2, 4, 8, 14, 30). We captured single cells with the 10x Genomics Chromium single-cell platform followed by quality control tests, and retained 5,2575 cells for subsequent transcriptomic analysis. Additionally, we assessed the feasibility of our system to model specific aspects of pregestational diabetes-induced congenital heart disease (PGD) in vitro. To simulate PGD, we modified our differentiation protocols to replicate the glucose levels associated with maternal hyperglycemia (defined as a random plasma glucose level of = 200 mg/dL) and performed scRNA-seq on populations differentiated under both PGD and control conditions.