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

Nature Protocols

Volumn 5, Issue 7, 2010, Pages 1265-1272

  Bernas, Michael J ^a   Cardoso, Filipa L ^b   Daley, Sarah K ^a   Weinand, Martin E ^a   Campos, Alexandre R ^c   Ferreira, António J Gonçalves ^c   Hoying, James B ^d   Witte, Marlys H ^a   Brites, Dora ^b   Persidsky, Yuri ^e   Ramirez, Servio H ^e   Brito, Maria A ^b  

^a UNIVERSITY OF ARIZONA COLLEGE OF MEDICINE   (United States)

^b UNIVERSITY OF LISBON   (Portugal)

^c HOSPITAL DE SANTA MARIA   (Portugal)

^d Cardiovascular Innovation Institute   (United States)

^e TEMPLE UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CAVEOLIN 1; CLAUDIN 5; GLUCOSE TRANSPORTER 1; GLYCOPROTEIN P; OCCLUDIN; PROTEIN ZO1; PROTEIN ZO2; VON WILLEBRAND FACTOR; CAV1 PROTEIN, HUMAN; COLLAGEN TYPE 1; MULTIDRUG RESISTANCE PROTEIN; SLC2A1 PROTEIN, HUMAN;

ARTICLE; BLOOD BRAIN BARRIER; CELL CULTURE; CELL EXPANSION; CELL ISOLATION; CLINICAL PROTOCOL; CONTROLLED STUDY; CULTURE TECHNIQUE; ENDOTHELIUM CELL; GENE EXPRESSION PROFILING; HUMAN; HUMAN CELL; HUMAN TISSUE; IMMUNOCYTOCHEMISTRY; IN VITRO STUDY; MATHEMATICAL MODEL; PERICYTE; PRIORITY JOURNAL; PROTEIN EXPRESSION; REPRODUCIBILITY; VALIDATION PROCESS; BIOLOGICAL MODEL; CYTOLOGY; HIPPOCAMPUS; IMPEDANCE; METABOLISM; PHYSIOLOGY; TEMPORAL LOBE; TIGHT JUNCTION;

BLOOD-BRAIN BARRIER; CAVEOLIN 1; CELL CULTURE TECHNIQUES; CELLS, CULTURED; COLLAGEN TYPE I; ELECTRIC IMPEDANCE; ENDOTHELIAL CELLS; GLUCOSE TRANSPORTER TYPE 1; HIPPOCAMPUS; HUMANS; MODELS, BIOLOGICAL; P-GLYCOPROTEIN; PERICYTES; TEMPORAL LOBE; TIGHT JUNCTIONS; VON WILLEBRAND FACTOR;

EID: 77954288447     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2010.76     Document Type: Article

References (40)

1. Persidsky, Y., Ramirez, S.H., Haorah, J. & Kanmogne, G.D. Blood-brain barrier: structural components and function under physiologic and pathologic conditions. J. Neuroimmune Pharmacol. 1, 223-236 (2006).
2. Weiss, N., Miller, F., Cazaubon, S. & Couraud, P.O. The blood-brain barrier in brain homeostasis and neurological diseases. Biochim. Biophys. Acta 1788, 842-857 (2009).
3. Zlokovic, B.V. The blood-brain barrier in health and chronic neurodegenerative disorders. Neuron 57, 178-201 (2008).
4. Ehrlich, P. Das Sauerstoffbedürfnis des Organismus, Eine Farbenanalytische Studie. Hirschwald, Berlin, (1885).
5. Joó, F. & Karnushina, I. A procedure for the isolation of capillaries from rat brain. Cytobios 8, 41-48 (1973).
6. Deli, M.A., Abrahám, C.S., Kataoka, Y. & Niwa, M. Permeability studies on in vitro blood-brain barrier models: physiology, pathology, and pharmacology. Cell Mol. Neurobiol. 25, 59-127 (2005).
7. Bowman, C. et al. Cystic hygroma reconsidered: hamartoma or neoplasm? Primary culture of an endothelial cell line from a massive cervicomediastinal cystic hygroma with bony lymphangiomatosis. Lymphology 17, 15-22 (1984).
8. Way, D. et al. Lymphatic endothelial cell line (CH3) from a recurrent retroperitoneal lymphangioma. In Vitro 23, 647-652 (1987).
9. Bowman, P.D. et al. Primary culture of capillary endothelium from rat brain. In Vitro 17, 353-362 (1981).
10. Bowman, P.D., Ennis, S.R., Rarey, K.E., Betz, A.L. & Goldstein, G.W. Brain microvessel endothelial cells in tissue culture: a model for study of blood-brain barrier permeability. Ann. Neurol. 14, 396-402 (1983).
11. Stins, M.F., Gilles, F. & Kim, K.S. Selective expression of adhesion molecules on human brain microvascular endothelial cells. J. Neuroimmunol. 76, 81-90 (1997).
12. Dorovini-Zis, K., Prameya, R. & Bowman, P.D. Culture and characterization of microvascular endothelial cells derived from human brain. Lab. Invest. 64, 425-436 (1991).
13. Dombrowski, S.M. et al. Overexpression of multiple drug resistance genes in endothelial cells from patients with refractory epilepsy. Epilepsia 42, 1501-1506 (2001).
14. Weksler, B.B. et al. Blood-brain barrier-specifc properties of a human adult brain endothelial cell line. FASEB J. 19, 1872-1874 (2005).
15. Hughes, C.C.W. & Lantos, P.L. Brain capillary endothelial cells in vitro lack surface IgG Fc receptors. Neurosci. Lett. 68, 100-106 (1986).
16. Hiu, T. et al. Tissue plasminogen activator enhances the hypoxia/reoxygenation-induced impairment of the blood-brain barrier in a primary culture of rat brain endothelial cells. Cell Mol. Neurobiol. 28, 1139-1146 (2008).
17. Coisne, C. et al. Mouse syngenic in vitro blood-brain barrier model: a new tool to examine infammatory events in cerebral endothelium. Lab. Invest. 85, 734-746 (2005).
18. Malina, K.C., Cooper, I. & Teichberg, V.I. Closing the gap between the in-vivo and in-vitro blood-brain barrier tightness. Brain Res. 1284, 12-21 (2009).
19. Colgan, O.C. et al. Infuence of basolateral condition on the regulation of brain microvascular endothelial tight junction properties and barrier function. Brain Res. 1193, 84-92 (2008).
20. Parkinson, F.E. & Hacking, C. Pericyte abundance affects sucrose permeability in cultures of rat brain microvascular endothelial cells. Brain Res. 1049, 8-14 (2005).
21. Perrière, N. et al. Puromycin-based purifcation of rat brain capillary endothelial cell cultures. Effect on the expression of blood-brain barrier-specifc properties. J. Neurochem. 93, 279-289 (2005).
22. Poller, B. et al. The human brain endothelial cell line hCMEC/D3 as a human blood-brain barrier model for drug transport studies. J. Neurochem. 107, 1358-1368 (2008).
23. Mkrtchyan, H. et al. Molecular cytogenetic characterization of the human cerebral microvessel endothelial cell line hCMEC/D3. Cytogenet. Genome Res. 126, 313-317 (2009).
24. Goldstein, G.W., Wolinsky, J.S. & Csejtey, J. Diamond, I. Isolation of metabolically active capillaries from rat brain. J. Neurochem. 25, 715-717 (1975).
25. Way, D.L. et al. In vitro models of angiotumotigenesis. Lymphology 27 (Suppl): 136-137 (1993).
26. Persidsky, Y. et al. A model for monocyte migration through the blood-brain barrier during HIV-1 encephalitis. J. Immunol. 158, 3499-3510 (1997).
27. Fiala, M. et al. TNF-α opens a paracellular route for HIV-1 invasion across the blood-brain barrier. Mol. Med. 3, 553-564 (1997).
28. Gan, X. et al. Cocaine enhances brain endothelial adhesion molecules and leukocyte migration. Clin. Immunol. 91, 68-76 (1999).
29. Persidsky, Y. et al. Microglial and astrocyte chemokines regulate monocyte migration through the blood-brain barrier in human immunodefciency virus-1 encephalitis. Am. J. Pathol. 155, 1599-1611 (1999).
30. Liu, N.Q. et al. Human immunodefciency virus type 1 enters brain microvascular endothelia by macropinocytosis dependent on lipid rafts and the mitogen-activated protein kinase signaling pathway. J. Virol. 76, 6689-6700 (2002).
31. Miebach, S. et al. Isolation and culture of microvascular endothelial cells from gliomas of different WHO grades. J. Neurooncol. 76, 39-48 (2006).
32. Shepherd, B.R., Chen, Y.S.H., Smith, C.M., Gruionu, G., Williams, S.K. & Hoying, J.B. Rapid perfusion and network remodeling in a microvascular construct after implantation. Arterioscler. Thromb. Vasc. Biol. 24, 898-904 (2004).
33. Fiala, M. et al. Cocaine increases human immunodefciency virus type 1 neuroinvasion through remodeling brain microvascular endothelial cells. J. Neurovirol. 11, 281-291 (2005).
34. Lossinsky, A.S. et al. The histopathology of Candida albicans invasion in neonatal rat tissues and in the human blood-brain barrier in culture revealed by light, scanning, transmission and immunoelectron microscopy. Histol. Histopathol. 21, 1029-1041 (2006).
35. Liu, J. et al. T cell independent mechanism for copolymer-1-induced neuroprotection. Eur. J. Immunol. 37, 3143-3154 (2007).
36. Haorah, J., Schall, K., Ramirez, S.H. & Persidsky, Y. Activation of protein tyrosine kinases and matrix metalloproteinases causes blood-brain barrier injury: novel mechanism for neurodegeneration associated with alcohol abuse. Glia 56, 78-88 (2008).
37. Ramirez, S.H. et al. Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) suppresses Rho GTPases in human brain microvascular endothelial cells and inhibits adhesion and transendothelial migration of HIV-1 infected monocytes. J. Immunol. 180, 1854-1865 (2008).
38. Zaghi, J. et al. Alzheimer disease macrophages shuttle amyloid-beta from neurons to vessels, contributing to amyloid angiopathy. Acta Neuropathol. 117, 111-124 (2009).
39. Ramirez, S.H. et al. Methamphetamine disrupts blood-brain barrier function by induction of oxidative stress in brain endothelial cells. J. Cereb. Blood Flow Metab. 29, 1933-1945 (2009).
40. Cardoso, FL. Establishment and characterization of a human model of the blood-brain barrier. Master Degree Thesis, New University of Lisbon 2009