title: Human Brain Microvascular Endothelial Cells Derived from the BC1 iPS Cell Line Exhibit a Blood-Brain Barrier Phenotype.
abstract: The endothelial cells that form capillaries in the brain are highly specialized, with tight junctions that minimize paracellular transport and an array of broad-spectrum efflux pumps that make drug delivery to the brain extremely challenging. One of the major limitations in blood-brain barrier research and the development of drugs to treat central nervous system diseases is the lack of appropriate cell lines. Recent reports indicate that the derivation of human brain microvascular endothelial cells (hBMECs) from human induced pluripotent stem cells (iPSCs) may provide a solution to this problem. Here we demonstrate the derivation of hBMECs extended to two new human iPSC lines: BC1 and GFP-labeled BC1. These hBMECs highly express adherens and tight junction proteins VE-cadherin, ZO-1, occludin, and claudin-5. The addition of retinoic acid upregulates VE-cadherin expression, and results in a significant increase in transendothelial electrical resistance to physiological values. The permeabilities of tacrine, rhodamine 123, and Lucifer yellow are similar to values obtained for MDCK cells. The efflux ratio for rhodamine 123 across hBMECs is in the range 2-4 indicating polarization of efflux transporters. Using the rod assay to assess cell organization in small vessels and capillaries, we show that hBMECs resist elongation with decreasing diameter but show progressive axial alignment. The derivation of hBMECs with a blood-brain barrier phenotype from the BC1 cell line highlights that the protocol is robust. The expression of GFP in hBMECs derived from the BC1-GFP cell line provides an important new resource for BBB research.
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Institute of Nanobiotechnology, 100 Croft Hall, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.
authors.forename: Moriah E
authors.initials: ME
authors.lastname: Katt
authors.affiliation: Institute of Nanobiotechnology, 100 Croft Hall, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Department of Biomedical Engineering, Johns Hopkins University, 720 Rutland Avenue, Baltimore, MD, 21205, United States of America.
authors.forename: Zinnia S
authors.initials: ZS
authors.lastname: Xu
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Institute of Nanobiotechnology, 100 Croft Hall, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.
authors.forename: Sharon
authors.initials: S
authors.lastname: Gerecht
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Institute of Nanobiotechnology, 100 Croft Hall, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.|Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, United States of America.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Searson
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D001812:Blood-Brain Barrier / Q000378:metabolism / Q000502:physiology*; D001921:Brain / Q000378:metabolism / Q000502:physiology*; D015820:Cadherins / Q000378:metabolism; D002196:Capillaries / Q000378:metabolism / Q000502:physiology; D002460:Cell Line; D062507:Claudin-5 / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D062793:Occludin / Q000378:metabolism; D010641:Phenotype; D020112:Rhodamine 123 / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism / Q000502:physiology; D015854:Up-Regulation / Q000502:physiology; D062826:Zonula Occludens-1 Protein / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D062507:Claudin-5; D062793:Occludin; D020112:Rhodamine 123; D062826:Zonula Occludens-1 Protein; D000099152:Cadherin 5; C569042:TJP1 protein, human