Fig. 1: Derivation of hPSC-derived vascular endothelial cells, smooth muscle cells, and pericytes.

A Schematic of directed differentiation of hPSCs to vascular endothelial cells (ECs), smooth muscle cells (SMCs), and pericytes (PCs) (Created using BioRender). B Bulk RNA sequencing was performed on hPSC-derived ECs, PCs, and SMCs. The normalized expression values of known EC, PC, or SMC marker genes were quantitated over three biological replicates. The values plotted represent scaled normalized expression values for each gene across samples (C) (left) Principal component analysis (PCA) on bulk-RNA sequencing data from hPSCs, hPSC-ECs, hPSC-PCs, and hPSC-SMCs and primary endothelial cells (HUVECs), primary pericytes, and primary bronchial smooth muscle cells (BSMCs). (right) Mural cell-only PCA analysis (Primary pericytes, BSMCs, hPSC-PCs, and hPSC-SMCs) (D) Expression of PECAM1 and VE-Cadherin was quantitated by flow cytometry on hPSCs or on two independent differentiations of ECs. Data points on the bar graph represent values from two independent differentiations. Expression of VE-Cadherin, VWF, and PECAM1 in hPSC-derived ECs was observed by immunofluorescence. E Expression of NG2 was quantitated by flow cytometry on hPSCs or on two independent differentiations of PCs. Data points on bar graph represent values from two independent differentiations. Expression of PDGFR-B, and PDGFR-A was quantitated by flow cytometry on HEK293 cells, human dermal fibroblast (hDF) and two independent PC differentiation. Expression of SMA, PDGFR-B, and NG2 in hPSC derived PCs was observed by immunofluorescence. F Expression of PDGFR-B or SMA was quantitated by flow cytometry on hPSCs or on two independent differentiations of SMCs. Data points on the bar graph represent values from two independent differentiations. Expression of SMA, PDGFR-B, and NG2 in hPSC-derived SMCs was observed by immunofluorescence. Scale bar = 50 µm for all immunofluorescence images.