Engineering of human brain organoids with a functional vascular-like system

Abstract Here, we engineered human embryonic stem cells (hESCs) to ectopically express human ETS variant 2 (ETV2) to create in vitro vasculature in cortical organoids (hCOs), namely vhCOs (vascularized hCOs). ETV2-expressing cells in hCOs contributed to forming a complex vascular-like network in hCOs. This protocol describes hESC maintenance, the generation of inducible ETV2 expressing hESCs, and the generation of cortical organoid with functional vascular-like network.