Hyperoxia impairs iPSC derived endothelial cells and drives an atherosclerosis-like transcriptional phenotype

Induced pluripotent stem cells (iPSCs) directed to endothelial identity (iPSC-ECs) lose expression of key identity markers under standard in vitro conditions, limiting their clinical applications. We examined iPSC-ECs at late passage (>2 weeks) under hyperoxic (21%)conditions with single cell RNA sequencing Y6 pluripotent stem cells were directed to endothelial identity by treatment with CHIR99021, a GSK3 inhibitor, as well as BMP4 for 3 days, followed by VEGFa and forskolin treatment for 2 days according to the protocol described by Cowen et al 2014, Nature Cell Biology DOI 10.1038/ncb3205. Then, cells were purified using CD144 magnetic beads, and were subsequently cultured for two weeks at atmospheric oxygen level (21%) in a conventional cell culture incubator before harvest and analysis by scRNAseq