Derivation of blood-brain barrier endothelial cells from human pluripotent stem cells

This 4-step protocol describes a method for differentiation of brain microvascular endothelial cells (BMECs) from human embryonic stem cells (hESCs) by inducing simultaneous endothelial and neural cell differentiation, followed by purification of the BMECs on a selective matrix. An additional step of co-culturing with astrocyte cells can be performed for further estimation of the blood-brain barrier (BBB) properties of the differentiated cells. The BMECs express BBB markers, respond to astrocytic cues, and have barrier and transport properties similar to those of primary BMECs.