RNA sequencing of hiPSC-derived endothelial cells via doxycycline-induced ETV2 transcription factor overexpression

Endothelial cells are known to express tissue-specific phenotypes to support the organ functionality due to the residing tissue microenvironment niche. The objective of this study is to compare the transcriptomic profiling of the generated endothelial cells (called iETV2-ECs) in neuro maintenance medium (NMM) with endothelial cells derived from primary human brain and other organs, as well as human induced pluripotent stem cells. The iETV2-ECs were generated from human induced pluripotent stem cells (hiPSC) that were genome engineered with a doxycycline-inducible overexpression cassette for the master regulator of endothelial differentiation (i.e. ETV2). The cells were treated with doxycycline for 8 days in the NMM supplemented with bFGF, FBS and endothelial cell growth supplements and subsequently maintained in NMM up to 21 days. Cells were harvested on day 8, 14 and 21 days for total RNA extraction, and their transcriptomic profiles were analyzed by bulk RNA sequencing technique. P ublicly-available RNA sequencing datasets of different endothelial cell types were integrated into the iETV2-EC data set for differentially expressed genes analysis. The ultimate goal of this project is to develop vascularized 3D neural model to study human brain vascularization and disease modelling. Overall design: The goal of this study is to analyze the transcriptomic profiling of the generated endothelial cells (iETV2-Ecs) in neuro maintenance medium. RNA profiles of endothelial cells at day 8, 14 and 21 days. Three independent samples for each time points (n=3)