Establishment of an blood-brain/tumor barrier modal in vitro by coculturing brain endothelial cells,pericytes and U251
10.3969/j.issn.1671-8348.2014.20.025
- VernacularTitle:应用U251胶质瘤细胞、脑内皮细胞和周细胞体外构建血瘤／脑屏障模型的研究
- Author:
Haiyang YANG
;
Li CAI
;
Jiang LONG
;
Jihong HE
- Publication Type:Journal Article
- Keywords:
blood-brain barrier;
endothelial cells;
pericytes;
U251 glioma cells
- From:
Chongqing Medicine
2014;(20):2613-2616,2619
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a stable in vitro model of blood-brain/tumor barrier (BBB/BTB) simulating in vivo state u-sing the human brain endothelial cells(hBMEC) ,brain pericytes (PC) and U251 glioma cells(U251) .Methods An in vitro model of BBB/BTB was constructed using Transwell inserts(pore size 1 .0 μm)coculture while three cell subculture reaches a certain num-ber .Its barrier function was evaluated by the 4-hour leakage test ,inverted microscope to observe morphological changes ,horseradish peroxidase permeability test and tight junction protein Claudin-5 and Occludin identification expression observed by immunofluores-cence technology .Results hBM EC monolayers demonstrated a typical cobblestone-like appearance ,Non-contact spindle cocultured monolayer growth appeared its uniqueswirl poolshape ,fusiform morphology ;the pericytes displayed irregular shape and overlap-ping grow ;U251 showed typical tumor cell growth .Fluid leakage test were interviewing a certain level difference in three different ways to build models ;immnocytochemical staining showed a continuous and dense tight junction formed between the endothelial cells but intensity ranging ;horseradish peroxidase permeability of three different models showed that pure hBMEC (43 .490 ± 3 .572)% ,hBMEC+U251(36 .540 ± 1 .475)% ,hBMEC+ PC+U251(26 .460 ± 2 .372)% was significantly lower ,the difference was statistically significant(t=19 .330 ,P<0 .01) .Conclusion hBMEC+ PC+ U251 cocultured the morphology ,tructure and barrier function have more advantages than pure hBMEC and hBMEC+U251 ,have the basic characteristics and function of BTB/BBB ,and they have provided a new tool reseach function ,regulation mechanisms and drug screening the BTB/BBB in future .
