title: Validation of in vitro cell culture models of the blood-brain barrier: tightness characterization of two promising cell lines.
abstract: In the course of the validation of blood-brain barrier in vitro models the aim of this work was to characterize two promising continuous cell lines with regard to their tightness properties. PBMEC/C1-2 and ECV304 cells were cultured in several media with different compositions on Transwell inserts. Inducibility and functionality of tightness were investigated by transendothelial electrical resistance (TEER) and by transport studies with transcellular marker diazepam, glycine antagonist Bu101 and paracellular marker APTS-dextran. Inducibility, expression and localization of tight junctional proteins were assessed by western blotting and immunofluorescence microscopy. Presence of factors derived from glioma cell line C6 resulted in increased TEER in both cell lines. Comparison to APTS-dextran data across Caco-2 layers emphasized that correlations between permeability of the paracellular marker and TEER are dependent on each investigated cell line and the corresponding growth medium. Presence and inducibility of tight junctional proteins ZO-1 and Occludin were proven for ECV304 layers. Cell line ECV304 seemed to be suitable for TEER dependent transport studies, whereas PBMEC/C1-2 showed higher potential for P-gP studies.
authors.affiliation: Department of Medicinal Chemistry, University of Vienna, Pharmacy Center, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Winfried
authors.initials: W
authors.lastname: Neuhaus
authors.forename: Verena E
authors.initials: VE
authors.lastname: Plattner
authors.forename: Michael
authors.initials: M
authors.lastname: Wirth
authors.forename: Bettina
authors.initials: B
authors.lastname: Germann
authors.forename: Bodo
authors.initials: B
authors.lastname: Lachmann
authors.forename: Franz
authors.initials: F
authors.lastname: Gabor
authors.forename: Christian R
authors.initials: CR
authors.lastname: Noe
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier; D015153:Blotting, Western; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D003911:Dextrans / Q000493:pharmacokinetics; D003975:Diazepam / Q000493:pharmacokinetics; D066298:In Vitro Techniques; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D051381:Rats; D013056:Spectrophotometry, Ultraviolet
chemical_list: D003470:Culture Media; D003911:Dextrans; D003975:Diazepam