Establishment of an in vitro blood-brain barrier model by co-culturing rat brain microvascular endothelial cells,pericytes and astrocytes
10.3969/j.issn.1001-1978.2015.05.027
- VernacularTitle:大鼠脑微血管内皮细胞与周细胞、星形胶质细胞共培养建立体外血脑屏障模型
- Author:
Yufeng ZHA
;
Xiaozhong FU
;
Shun ZHANG
;
Min LUO
;
Yu OU
;
Yongxi DONG
;
Aimin WANG
;
Yonglin WANG
- Publication Type:Journal Article
- Keywords:
primitive culture;
brain microvascular endothelial cells;
pericytes;
astrocytes;
blood-brain barrier;
morphology;
immunocytochemistry
- From:
Chinese Pharmacological Bulletin
2015;(5):730-735
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish in vitro blood-brain barrier (BBB) model with characteristics of simulation of in vivo BBB by primi-tive co-culture of brain-microvessel endothelial cells (BMECs) with brain-microvessel pericytes (BMPC)and astrocytes (AS). Methods BMECs,BMPC and AS from SD rats were primitively isolated,purified and cultured,and then primitive culture cells were identified by cellular morphological and immunocytochemi-cal staining methods.Five types of in vitro BBB models were es-tablished by using Millicell culture insert (pore diameter 0.4μm)and their barrier functions were evaluated by detection of transendothelial electrical resistance (TEER),permeability of sodium fluorescent (Na-FLU ),expression of alkaline phospha-tase (AKP)and γ-glutamyl transpeptidase (γ-GT1 ),and simi-larity of permeation amount for positive drugs in vitro and in vivo BBB conditions.Results Primitive culture of BMECs presented typical pebbles-like structure,BMPC presented larger soma with branching property,AS presented slender synapse and shallower cytoplasm.Moreover,immunocytochemical staining results iden-tified primitive cells were targeted cells.TEER value for co-cul-ture of BMECs,BMPC and AS reached (478 ±25 )Ω· cm2 , permeability coefficients (Papp )value of Na-FLU was [(8.23 ± 0.78) ×10 -6 ]cm·s-1 ,expression of AKP and γ-GT1 were (6.90 ±0.27 )King unit · g-1 Pro and (4.39 ±0.32 )μg · g-1 Pro respectively.Moreover,good correlation could be found in Papp for positive controls in vitro and in vivo BBB models (R2=0.92).Conclusion The established in vitro BBB model by using primitive co-culture of BMECs with BMPC and AS posses-ses in vivo BBB properties in cell morphology,structures and barrier functions,and can be used as a powerful tool for studying physiology,pathology of BBB and screening candidate com-pounds.
