This study describes a two-step, chemically defined, xeno-free method for generating valve endothelial-like cells (VELs) from human pluripotent stem cells (hPSCs). hPSCs were first differentiated into KDR+/ISL1+ cardiac progenitors (CPCs), then into VELs via an intermediate endocardial cushion cell (ECC) stage. TGFβ1 and BMP4 were key in specifying VEC fate by activating NOTCH/WNT signaling and targets like ATF3 and KLF transcription factors. hPSC-derived VELs, when seeded onto decellularized porcine aortic valve (DCV) scaffolds, showed superior proliferation and clonogenicity compared to primary VECs and HAECs. This method provides efficient generation of hPSC-derived valvular cells for potential use in valve organoids or tissue-engineered heart valves.

LinXi Cheng, MingHui Xie, WeiHua Qiao, Yu Song, YanYong Zhang, YingChao Geng, WeiLin Xu, Lin Wang, Zheng Wang, Kai Huang, NianGuo Dong, YuHua Sun