Porcine brain microvessel endothelial cells as an in vitro model to predict in vivo blood-brain barrier permeability.
The objective of the study was to establish primary cultured porcine brain microvessel endothelial cells (PBMECs) as an in vitro model to predict the blood-brain barrier (BBB) permeability in vivo. The intercellular tight junction formation of PBMECs was examined by electron microscopy and measured by transendothelial electrical resistance (TEER). The mRNA expression of several BBB transporters in PBMECs was determined by reverse transcriptionpolymerase chain reaction analysis. The in vitro permeability of 16 structurally diverse compounds, representing a range of passive diffusion and transporter-mediated mechanisms of brain penetration, was determined in PBMECs. Except for the perfusion flow rate marker diazepam, the BBB permeability of these compounds was determined either in our laboratory or as reported in literature using in situ brain perfusion technique in rats. Results in the present study showed that PBMECs had a high endothelium homogeneity, an mRNA expression of several BBB transporters, and high TEER values. Culturing with rat astrocyte-conditioned medium increased the TEER of PBMECs, but had no effect on the permeability of sucrose, a paracellular diffusion marker. The PBMEC permeability of lipophilic compounds measured under stirred conditions was greatly increased compared with that measured under unstirred conditions. The PBMEC permeability of the 15 test compounds, determined under the optimized study conditions, correlated with the in situ BBB permeability with an r2 of 0.60. Removal of the three system L substrates increased the r2 to 0.89. In conclusion, the present PBMEC model may be used to predict or rank the in vivo BBB permeability of new chemical entities in a drug discovery setting.
Zhang
Yan
Y
Department of Pharmacokinetics, Dynamics & Metabolism, Global Research & Development, Pfizer Inc, Ann Arbor, MI 48105, USA.
Li
Cheryl S W
CS
Ye
Yuyang
Y
Johnson
Kjell
K
Poe
Julie
J
Johnson
Shannon
S
Bobrowski
Walter
W
Garrido
Rosario
R
Madhu
Cherukury
C
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001812:Blood-Brain Barrier / Q000378:metabolism*; D001921:Brain / Q000098:blood supply / Q000166:cytology; D002199:Capillary Permeability; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D066298:In Vitro Techniques; D008297:Male; D046529:Microscopy, Electron, Transmission; D008858:Microscopy, Phase-Contrast; D008954:Models, Biological; D010477:Perfusion; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011237:Predictive Value of Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D013552:Swine
D004364:Pharmaceutical Preparations