RNA-sequencing of human pluripotent stem-cell derived endothelial cells under control and Wnt-activating conditions

Endothelial cells (ECs) in the central nervous system (CNS) acquire specialized barrier properties in response to extrinsic signals, with Wnt/β-catenin signaling coordinating multiple aspects of endothelial barrier function. We used human pluripotent stem cell (hPSC)-derived endothelial progenitor cells to profile the EC response to Wnt activation using multiple strategies, including the small molecule GSK-3 inhibitor CHIR 99021, and the CNS-derived ligands Wnt7a and Wnt7b. 20 total samples were analyzed, 18 endothelial cell (EC) samples and 2 smooth muscle-like cell (SMLC) samples. All cells were differentiated from the human induced pluripotent stem cell line iPS(IMR90)-4. Four independent differentiations were performed (denoted 2, 3, 5, and 6). At Passage 1, ECs treated with DMSO (vehicle control), 4 uM CHIR 99021 (CHIR), or 50 ng/ml each Wnt7a and Wnt7b (WNT) were analyzed from differentiations 2, 3, 5, and 6. At Passage 3, ECs treated with DMSO or CHIR were analyzed from differentiations 2, 3, and 6. At Passage 1, SMLCs treated with DMSO were analyzed from differentiations 3 and 6. For Passage 1 samples, fluoresence-activated cell sorting was used to isolate CD31+ (EC) and CD31- (SMLC) populations.