title: Differentiation and characterization of human pluripotent stem cell-derived brain microvascular endothelial cells.
abstract: The blood-brain barrier (BBB) is a critical component of the central nervous system (CNS) that regulates the flux of material between the blood and the brain. Because of its barrier properties, the BBB creates a bottleneck to CNS drug delivery. Human in vitro BBB models offer a potential tool to screen pharmaceutical libraries for CNS penetration as well as for BBB modulators in development and disease, yet primary and immortalized models respectively lack scalability and robust phenotypes. Recently, in vitro BBB models derived from human pluripotent stem cells (hPSCs) have helped overcome these challenges by providing a scalable and renewable source of human brain microvascular endothelial cells (BMECs). We have demonstrated that hPSC-derived BMECs exhibit robust structural and functional characteristics reminiscent of the in vivo BBB. Here, we provide a detailed description of the methods required to differentiate and functionally characterize hPSC-derived BMECs to facilitate their widespread use in downstream applications.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Stebbins
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States.
authors.forename: Hannah K
authors.initials: HK
authors.lastname: Wilson
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States.
authors.forename: Scott G
authors.initials: SG
authors.lastname: Canfield
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States.
authors.forename: Tongcheng
authors.initials: T
authors.lastname: Qian
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States. Electronic address: sppalecek@wisc.edu.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Palecek
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, WI 53706, United States. Electronic address: eshusta@wisc.edu.
authors.forename: Eric V
authors.initials: EV
authors.lastname: Shusta
mesh_terms: D001812:Blood-Brain Barrier / Q000166:cytology; D001921:Brain / Q000098:blood supply; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D055806:Microvessels / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000502:physiology*
keywords: Blood–brain barrier; Brain microvascular endothelial cells; Differentiation; Human pluripotent stem cells