title: Murine in vitro model of the blood-brain barrier for evaluating drug transport.
abstract: In vitro blood-brain barrier (BBB) models help predict brain uptake of potential central nervous system drug candidates. Current in vitro models are composed of brain microvascular endothelial cells (BMEC) that are isolated from rat, bovine, or porcine. However, most in vivo studies on drug transport through the BBB are performed in small laboratory animals, specially mouse and thus murine in vitro BBB models serve as better surrogates to correlate with these studies. Here we describe the functional characterization of a reproducible in vitro model composed of murine BMEC co-cultured with rat primary astrocytes in the presence of biochemical inducing agents. The co-cultures presented high TEER and low sodium fluorescein permeability. Expression of specific BBB tight junction proteins (occludin, claudin-5, ZO-1) and the functionality of transporters (Pgp, GLUT1) were detected by immunocytochemistry and Western blotting. These results indicated a 2.5-fold increase in the expression levels of these proteins in the presence of astrocytes. In addition, a high correlation coefficient (0.98) was obtained between the permeability of a series of hydrophobic and hydrophilic drugs and their corresponding in vivo values. These results together establish the utility of this murine model for future drug transport, pathological, and pharmacological characterizations of the BBB.
authors.affiliation: Department of Biomedical Engineering, Cornell University, Ithaca, NY, USA.
authors.forename: Gilda
authors.initials: G
authors.lastname: Shayan
authors.forename: Yong Seok
authors.initials: YS
authors.lastname: Choi
authors.forename: Eric V
authors.initials: EV
authors.lastname: Shusta
authors.forename: Michael L
authors.initials: ML
authors.lastname: Shuler
authors.forename: Kelvin H
authors.initials: KH
authors.lastname: Lee
mesh_terms: D000818:Animals; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D002491:Central Nervous System Agents / Q000493:pharmacokinetics*; D018920:Coculture Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D010539:Permeability; D051381:Rats; D019108:Tight Junctions / Q000378:metabolism
chemical_list: D002491:Central Nervous System Agents; D008565:Membrane Proteins