Contributions of BMPR2 Mutations and Extrinsic Factors to Cellular Phenotypes of Pulmonary Arterial Hypertension Revealed by Induced Pluripotent Stem Cell …

MethodsUsing clustered regularly interspaced short palindromic repeats–Cas9–mediated homologous recombination in a wild-type iPSC line, two isogenic sublines carrying either a known causal BMPR2 mutation (W9X; referred to as C2 W9X+/−) or a deletion of exon 1 (C2 ΔExon1) were generated. Serum-free, chemically defined iPSC differentiation protocols were used to generate iPSC-derived SMCs (iPSC-SMCs) and iPSC-ECs. This was achieved by differentiating iPSCs into iPSC-SMCs via a lateral plate mesoderm, paraxial mesoderm, or neural ectoderm lineage followed by 12 days in TGF-β1 (transforming growth factor β 1) and PDGF-BB (platelet-derived growth factor BB)±BMP4 (bone morphogenetic protein 4)(Figure 1A)(3), and into ECs via FGF-2 (fibroblast growth factor 2)–induced, BMP4-induced, and LY294002-induced mesoderm followed by FGF-2 and VEGF (vascular endothelial growth factor …