title: Astrocytic contributions to blood-brain barrier (BBB) formation by endothelial cells: a possible use of aortic endothelial cell for in vitro BBB model.
abstract: Astrocytic contribution of endothelial cell monolayer permeability was examined in two blood-brain barrier (BBB) models, using the coculture in a double chamber system: rat astrocytes and bovine aortic endothelial cells (BAECs) or bovine brain endothelial cells (BBECs). In system 1, where astrocytes were separated from endothelial cells, a 40% reduction in L-glucose permeability of the BBEC monolayer, but not the BAEC monolayer, was observed by cocultivation with astrocytes. Although several passages of BBEC in culture elicited morphological transformation from spindle-shapes to cobblestone-like features, the passaged BBECs remained responsive to astrocytes in coculture in system 1 (37% reduction of the L-glucose permeability). By contrast, in system 2, where respective endothelial cells and astrocytes layered on the upper and lower surfaces of a membrane, the permeability of both BAEC and BBEC monolayers was reduced by cocultivation with astrocytes (75% reduction for BAEC and 40% reduction for BBEC). BAECs in this contiguous coculture (system 2) with astrocytes showed numerous tight junction-like structures characteristic of the BBB in vivo. These results suggest that primary cultured BBECs, which had been primed by astrocytes in vivo, retain a higher sensitivity to astrocytes possibly through an astrocytic soluble factor (s) to exhibit BBB-specific phenotypes, and that even BAEC from extra-neural tissues, when cultured with astrocytes in close proximity in vitro, may acquire the similar phenotypes and serve for an extensive use of BBB model in vitro.
authors.affiliation: Department of Bioregulation Research, Nagoya City University Medical School, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Isobe
authors.forename: T
authors.initials: T
authors.lastname: Watanabe
authors.forename: T
authors.initials: T
authors.lastname: Yotsuyanagi
authors.forename: N
authors.initials: N
authors.lastname: Hazemoto
authors.forename: K
authors.initials: K
authors.lastname: Yamagata
authors.forename: T
authors.initials: T
authors.lastname: Ueki
authors.forename: K
authors.initials: K
authors.lastname: Nakanishi
authors.forename: K
authors.initials: K
authors.lastname: Asai
authors.forename: T
authors.initials: T
authors.lastname: Kato
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000502:physiology; D001253:Astrocytes / Q000502:physiology*; D001812:Blood-Brain Barrier / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005947:Glucose / Q000493:pharmacokinetics; D008854:Microscopy, Electron; D008954:Models, Biological; D010641:Phenotype; D051381:Rats
chemical_list: D005947:Glucose