[Establishment of an in vitro model of brain-blood barrier].
OBJECTIVE
To establish an in vitro model of brain-blood barrier (BBB) using cultured mouse brain microvascular endothelial cells (BMVEC).


METHODS
Mouse BMVEC were seeded on micro-pore membrane of gelatin-coated cell culture insert and cultured to confluence. The establishment of BBB was preliminary judged by a 4 h water-leaking test. The tight junctions between BMVEC were demonstrated by scanning and transmission electron microscope. The transendothelial electrical resistance(TEER) over BMVEC was measured. The permeability of Horseradish peroxidase (HRP) through the BBB was analyzed and the effect of RMP-7 on permeability of the BBB was investigated.


RESULTS
The 4 h water-leaking test became positive when BMVEC were cultured to confluence. By scanning and transmission electron microscope, the tight junctions were demonstrated on confluent BMVEC. The TEER over BMVEC monolayer increased 3.2 and 7.68 times and the permeability rates for HRP were 13.4% and 6.7% respectively, as compared with sub-confluent BMVEC and human umbilical vein endothelial cell monolayer(HUVEC). The HRP permeability rate in the model of BBB increased 2.7 times after treatment with RMP-7.


CONCLUSIONS
The established in vitro model of BBB has basic characteristics of BBB in vivo, and is suitable for central nervous system (CNS) drug research over BBB.
Xie
Ying
Y
Department of Pharmaceutics, Peking University School of Pharmaceutical Sciences, Beijing 100083, China.
Ye
Li-ya
LY
Zhang
Xiao-bin
XB
Hou
Xin-pu
XP
Lou
Jin-ning
JN
D000818:Animals; D001812:Blood-Brain Barrier; D001921:Brain / Q000098:blood supply*; D002196:Capillaries / Q000166:cytology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000648:ultrastructure; D051379:Mice; D008808:Mice, Inbred CBA; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission