[Immortalized mouse brain endothelial cell line Bend.3 displays the comparative barrier characteristics as the primary brain microvascular endothelial cells].
OBJECTIVE
The purpose of this study was to assess weather the immortalized mouse brain endothelial cell line Bend.3 displays the comparative barrier characteristics as the primary brain microvascular endothelial cells (BEMC).


METHODS
Immortalized mouse brain endothelial cell line, Bend.3 cells were cultured in transwell inserts and their restrictive characteristics were assessed by transendothelial electrical resistance (TEER) and horseradish peroxidase (HRP) permeability assays. Western blot and direct fluorescent staining methods were used to detect the tight junction protein expression and F-actin distribution.


RESULTS
The TEER in Bend.3 cells increased with the prolonged culture time and increased to 82.3+/-6.0 Omega cm2 10 days after culture, which was significantly higher than that 3 days after culture (37.3+/-3.1 Omega cm2; P<0.05). There were significant differences in the permeability rates for HRP 3 and 10 days after culture (4.3+/-0.20)% vs (2.2+/-0.05)% (P<0.05). Western blot indicated high level expression of tight junction proteins occludin and ZO-1 in Bend.3 cells 10 days after culture. F-actin was visualized around the cell membrane and presented scrobiculate linear fluorescence 10 days after culture.


CONCLUSIONS
Bend.3 cells have similar barrier characteristics to BEMC, and their barrier function may reach to the best effect 10 days after culture.
He
Fang
F
Department of Pediatrics, Xiangya Hospital, Central South University, Changsha 410008, China.
Yin
Fei
F
Peng
Jing
J
Li
Kong-Zhao
KZ
Wu
Li-Wen
LW
Deng
Xiao-Lu
XL
D000199:Actins / Q000032:analysis; D000818:Animals; D001812:Blood-Brain Barrier; D002460:Cell Line; D017097:Electric Impedance; D042783:Endothelial Cells / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D010750:Phosphoproteins / Q000032:analysis; D062826:Zonula Occludens-1 Protein
D000199:Actins; D008565:Membrane Proteins; D010750:Phosphoproteins; C569043:Tjp1 protein, mouse; D062826:Zonula Occludens-1 Protein; D006735:Horseradish Peroxidase