Protocol for generation of endothelial cells from human-derived iPSCs and integration into mouse brain explants

The rapid and reliable generation of endothelial cells from induced pluripotent stem cells (iPSCs) is a keystone for biomedical research and regenerative medicine. Here, we present a protocol for human iPSC genetic engineering, ETS variant 2 (ETV2)-mediated endothelial differentiation, and subsequent injection in ex vivo mouse brain explants to study cellular interactions. By leveraging optimized techniques and tackling common technical problems, we provide a platform that enables physiologically relevant studies of vascular behavior, plasticity, and interactions within the neural microenvironment. For complete details on the use and execution of this protocol, please refer to Arce et al.¹.