Establishment of primary cultures of human brain microvascular endothelial cells to provide an in vitro cellular model of the blood-brain barrier

被引：164

作者：

Bernas, Michael J. ^{[2
]}

Cardoso, Filipa L. ^{[1
]}

Daley, Sarah K. ^{[2
]}

Weinand, Martin E. ^{[2
]}

Campos, Alexandre R. ^{[3
]}

Goncalves Ferreira, Antonio J. ^{[3
]}

Hoying, James B. ^{[4
]}

Witte, Marlys H. ^{[2
]}

Brites, Dora ^{[1
]}

Persidsky, Yuri ^{[5
]}

Ramirez, Servio H. ^{[5
]}

Brito, Maria A. ^{[1
]}

机构：

[1] Univ Lisbon, Fac Farm, Ctr Pathogenese Mol UBMBE iMed, P-1699 Lisbon, Portugal

[2] Univ Arizona, Dept Surg, Tucson, AZ USA

[3] Hosp Santa Maria, Serv Neurocirurgia, Lisbon, Portugal

[4] Cardiovasc Innovat Inst, Louisville, KY USA

[5] Temple Univ, Sch Med, Dept Pathol & Lab Med, Philadelphia, PA 19122 USA

来源：

NATURE PROTOCOLS | 2010年 / 5卷 / 07期

关键词：

RAT-BRAIN; ADHESION MOLECULES; MONOCYTE MIGRATION; LINE HCMEC/D3; PERMEABILITY; ENCEPHALITIS; CAPILLARIES; ACTIVATION; EXPRESSION; MECHANISM;

D O I：

10.1038/nprot.2010.76

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a method for generating primary cultures of human brain microvascular endothelial cells (HBMVECs). HBMVECs are derived from microvessels isolated from temporal tissue removed during operative treatment of epilepsy. The tissue is mechanically fragmented and size filtered using polyester meshes. The resulting microvessel fragments are placed onto type I collagen-coated flasks to allow HBMVECs to migrate and proliferate. The overall process takes less than 3 h and does not require specialized equipment or enzymatic processes. HBMVECs are typically cultured for approximately 1 month until confluent. Cultures are highly pure (similar to 97% endothelial cells; similar to 3% pericytes), are reproducible, and show characteristic brain endothelial markers (von Willebrand factor, glucose transporter-1) and robust expression of tight and adherens junction proteins as well as caveolin-1 and efflux protein P-glycoprotein. Monolayers of HBMVECs show characteristically high transendothelial electric resistance and have proven useful in multiple functional studies for in vitro modeling of the human blood-brain barrier.

引用

页码：1265 / 1272

页数：8

共 40 条

[1]

BOWMAN CA, 1984, LYMPHOLOGY, V17, P15

[2] BRAIN MICROVESSEL ENDOTHELIAL-CELLS IN TISSUE-CULTURE - A MODEL FOR STUDY OF BLOOD-BRAIN-BARRIER PERMEABILITY [J].

BOWMAN, PD ;

ENNIS, SR ;

RAREY, KE ;

BETZ, AL ;

GOLDSTEIN, GW .

ANNALS OF NEUROLOGY, 1983, 14 (04) :396-402

[3]

BOWMAN PD, 1981, IN VITRO CELL DEV B, V17, P353

[4]

CARDOSO FL, 2009, THESIS NEW U LISBON

[5] Mouse syngenic in vitro blood-brain barrier model:: a new tool to examine inflammatory events in cerebral endothelium [J].

Coisne, C ;

Dehouck, L ;

Faveeuw, C ;

Delplace, Y ;

Miller, F ;

Landry, C ;

Morissette, C ;

Fenart, L ;

Cecchelli, R ;

Tremblay, P ;

Dehouck, B .

LABORATORY INVESTIGATION, 2005, 85 (06) :734-746

[6] Influence of basolateral condition on the regulation of brain microvascular endothelial tight junction properties and barrier function [J].

Colgan, Olga C. ;

Collins, Nota T. ;

FeTguson, Gail ;

Murphy, Ronan P. ;

Bitney, Yvonne A. ;

Cahill, Paul A. ;

Cummins, Philip M. .

BRAIN RESEARCH, 2008, 1193 :84-92

[7] Permeability studies on in vitro blood-brain barrier models: Physiology, pathology, and pharmacology [J].

Deli, MA ;

Abraham, CS ;

Kataoka, Y ;

Niwa, M .

CELLULAR AND MOLECULAR NEUROBIOLOGY, 2005, 25 (01) :59-127

[8] Overexpression of multiple drug resistance genes in endothelial cells from patients with refractory epilepsy [J].

Dombrowski, SM ;

Desai, SY ;

Marroni, M ;

Cucullo, L ;

Goodrich, K ;

Bingaman, W ;

Mayberg, MR ;

Bengez, L ;

Janigro, D .

EPILEPSIA, 2001, 42 (12) :1501-1506

[9]

DOROVINIZIS K, 1991, LAB INVEST, V64, P425

[10]

Ehrlich P., 1885, SAUERSTOFFBEDURFNIS

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