Transcription Factor Driven Generation of Vascular Organoids from Induced Pluripotent Stem Cells

We present a streamlined method for generating vascularized organoids through transcription factor-mediated induction. Naïve (2iL) Mouse embryonic stem cells are lentivirally transduced with Etv2 and Nkx3.1, followed by clonal selection and conversion into epiblast-like cells (EpiLCs). This approach bypasses the need for exogenous growth factor supplementation and extracellular matrix embedding, resulting in the spontaneous formation of 3D vascular spheroids composed of integrated endothelial and mural cell networks. The resulting organoids exhibit functional vascular organization and provide a simple, reproducible platform for studying vascular development and tissue regeneration within a physiologically relevant 3D microenvironment.