A new blood–brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes

• Published in: Neurochemistry international Vol. 54; no. 3; pp. 253 - 263
• Main Authors: Nakagawa, Shinsuke, Deli, Mária A., Kawaguchi, Hiroko, Shimizudani, Takeshi, Shimono, Takanori, Kittel, Ágnes, Tanaka, Kunihiko, Niwa, Masami
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.03.2009; Elsevier
• Subjects: Animals; Astrocytes; Astrocytes - metabolism; Astrocytes - ultrastructure; ATP Binding Cassette Transporter, Sub-Family B - metabolism; ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism; Biological and medical sciences; Blood-Brain Barrier - metabolism; Blood-Brain Barrier - ultrastructure; Blood–brain barrier; Brain endothelial cells; Capillaries - metabolism; Capillaries - ultrastructure; Carrier Proteins - metabolism; Cell Membrane Permeability - drug effects; Cell Membrane Permeability - physiology; Cells, Cultured; Cerebral Arteries - metabolism; Cerebral Arteries - ultrastructure; Cerebral circulation. Blood-brain barrier. Choroid plexus. Cerebrospinal fluid. Circumventricular organ. Meninges; Claudin-5; Co-culture; Drug permeability; Endothelial Cells - metabolism; Endothelial Cells - ultrastructure; Fundamental and applied biological sciences. Psychology; In vitro BBB model (rat); Membrane Potentials - physiology; Membrane Proteins - metabolism; Microcirculation - physiology; Microscopy, Electron, Transmission; Models, Biological; Occludin; P-Glycoprotein; Pericytes; Pericytes - metabolism; Pericytes - ultrastructure; Phosphoproteins - metabolism; Rats; Rhodamine 123 - pharmacokinetics; Tight junction; Tight Junctions - metabolism; Tight Junctions - ultrastructure; Transendothelial electrical resistance; Vertebrates: nervous system and sense organs; Zonula Occludens-1 Protein; Astrocytes; Pericytes; In vitro BBB model (rat); Tight junction; Transendothelial electrical resistance; Co-culture; P-Glycoprotein; Drug permeability; Blood–brain barrier; Brain endothelial cells; Animal model; Pericyte; Endothelial cell; Rat; Neuroglia; Central nervous system; Cell junction; Blood brain barrier; Encephalon; Blood-brain barrier; Rodentia; Glycoprotein; Astrocyte; Permeability; In vitro; Vertebrata; Mammalia; Animal; Electric resistivity
• ISSN: 0197-0186; 1872-9754
• DOI: 10.1016/j.neuint.2008.12.002

Blood–brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells and neighbouring elements of the neurovascular unit. While pericytes are the cells situated closest to brain endothelial cells morphologically and share a common basement membrane, they have not been used in co-culture BBB models for testing drug permeability. We have developed and characterized a new syngeneic BBB model using primary cultures of the three main cell types of cerebral microvessels. The co-culture of endothelial cells, pericytes and astrocytes mimick the anatomical situation in vivo. In the presence of both pericytes and astrocytes rat brain endothelial cells expressed enhanced levels of tight junction (TJ) proteins occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. Further morphological evidence of the presence of interendothelial TJs was provided by electron microscopy. The transendothelial electrical resistance (TEER) of brain endothelial monolayers in triple co-culture, indicating the tightness of TJs reached 400 Ω cm 2 on average, while the endothelial permeability coefficients ( P e) for fluorescein was in the range of 3 × 10 −6 cm/s. Brain endothelial cells in the new model expressed glucose transporter-1, efflux transporters P-glycoprotein and multidrug resistance protein-1, and showed a polarized transport of rhodamine 123, a ligand for P-glycoprotein. To further characterize the model, drug permeability assays were performed using a set of 19 compounds with known in vivo BBB permeability. Good correlation ( R 2 = 0.89) was found between in vitro P e values obtained from measurements on the BBB model and in vivo BBB permeability data. The new BBB model, which is the first model to incorporate pericytes in a triple co-culture setting, can be a useful tool for research on BBB physiology and pathology and to test candidate compounds for centrally acting drugs.