title: Limitations of the hCMEC/D3 cell line as a model for Aβ clearance by the human blood-brain barrier.
abstract: Alzheimer's disease and cerebral amyloid angiopathy are characterized by accumulation of amyloid-β (Aβ) at the cerebrovasculature due to decreased clearance at the blood-brain barrier (BBB). However, the exact mechanism of Aβ clearance across this barrier has not been fully elucidated. The hCMEC/D3 cell line has been characterized as a valid model for the BBB. In this study we evaluated the use of this model to study Aβ clearance across the BBB, with an emphasis on brain-to-blood directional permeability. Barrier integrity of hCMEC/D3 monolayers was confirmed for large molecules in both the apical to basolateral and the reverse direction. However, permeability for smaller molecules was substantially higher, especially in basolateral to apical direction, and barrier formation for Aβ was completely absent in this direction. In addition, hCMEC/D3 cells failed to develop a high TEER, possibly caused by incomplete formation of tight junctions. We conclude that the hCMEC/D3 model has several limitations to study the cerebral clearance of Aβ. Therefore, the model needs further characterization before this cell system can be generally applied as a model to study cerebral Aβ clearance. © 2016 The Authors Journal of Neuroscience Research Published by Wiley Periodicals, Inc.
authors.affiliation: Radboud University Medical Center, Donders Institute for Brain, Cognition and Behaviour, Departments of Neurology and Laboratory Medicine, Radboud Alzheimer Centre, Nijmegen, The Netherlands.
authors.forename: Elisanne A L M
authors.initials: EALM
authors.lastname: Biemans
authors.affiliation: Radboud University Medical Center, Donders Institute for Brain, Cognition and Behaviour, Departments of Neurology and Laboratory Medicine, Radboud Alzheimer Centre, Nijmegen, The Netherlands.
authors.forename: Lieke
authors.initials: L
authors.lastname: Jäkel
authors.affiliation: Radboud University Medical Center, Department of Pathology, Nijmegen, The Netherlands.
authors.forename: Robert M W
authors.initials: RMW
authors.lastname: de Waal
authors.affiliation: Radboud University Medical Center, Donders Institute for Brain, Cognition and Behaviour, Departments of Neurology and Laboratory Medicine, Radboud Alzheimer Centre, Nijmegen, The Netherlands.
authors.forename: H Bea
authors.initials: HB
authors.lastname: Kuiperij
authors.affiliation: Radboud University Medical Center, Donders Institute for Brain, Cognition and Behaviour, Departments of Neurology and Laboratory Medicine, Radboud Alzheimer Centre, Nijmegen, The Netherlands.
authors.forename: Marcel M
authors.initials: MM
authors.lastname: Verbeek
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism*; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D019108:Tight Junctions / Q000378:metabolism*
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42)
keywords: amyloid-β; blood-brain barrier; clearance; hCMEC/D3; in vitro model