Blood-brain barrier (BBB) models are often used to investigate BBB function and screen brain-penetrating therapeutics, but it has been difficult to construct a human model that possesses an optimal BBB phenotype and is readily scalable. To address this challenge, we developed a human in vitro BBB model comprising brain microvascular endothelial cells (BMECs), pericytes, astrocytes and neurons derived from renewable cell sources. First, retinoic acid (RA) was used to substantially enhance BBB phenotypes in human pluripotent stem cell (hPSC)-derived BMECs, particularly through adherens junction, tight junction, and multidrug resistance protein regulation. RA-treated hPSC-derived BMECs were subsequently co-cultured with primary human brain pericytes and human astrocytes and neurons derived from human neural progenitor cells (NPCs) to yield a fully human BBB model that possessed significant tightness as measured by transendothelial electrical resistance (∼5,000 Ωxcm2). Overall, this scalable human BBB model may enable a wide range of neuroscience studies.
Bibliographical noteFunding Information:
We would like to thank the WiCell Research Institute for providing reagents and Dr. Clive Svendsen (Cedars-Sinai Medical Center, Los Angeles, CA) for providing the human NPCs used in this study7. E.S.L. was supported by a Chemistry Biology Interface training fellowship (T32 GM008505) and S.M.A. was supported by a National Science Foundation graduate fellowship. This work was funded in part by the US National Institutes of Health (NIH) grants AA020476 and NS083688 to E.V.S. and S.P.P.