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hPSCs were dissociated into single-cell suspensions using TrypLE Select (Thermo Fisher Scientific, Netherlands) and plated at a density of 3.5 10 6 cells/well in E8 medium (Thermo Fisher Scientific, Netherlands) supplemented with 10 M Y-27632 (Peprotech, USA) on low-adhesion, non-treated 6-well plates (Eppendorf, Germany) placed on an orbital shaker at 37 C, 110 rpm, under a 5% CO 2 atmosphere

a model of endothelial cell culture was used to mimic the environment of pulmonary hypertension, combining both chemical and physical stimuliNPs and cyclic stretch, respectively [220]