Immortalized Human Brain Endothelial Cells and Flow-Based Vascular Modeling: A Marriage of Convenience for Rational Neurovascular Studies

Journal of Cerebral Blood Flow and Metabolism - Tập 28 Số 2 - Trang 312-328 - 2008
Luca Cucullo1,2, Pierre‐Olivier Couraud3,4,5,6, Babette Weksler7,5, Ignacio A. Romero8, Mohammed Kamal Hossain1,2, Edward Rapp9, Damir Janigro10,1,2
1Department of Neurosurgery, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA
2Division of Cerebrovascular Research, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA
3CNRS UMR 8104, Paris, France
4Department of Cell Biology, Institut Cochin, Paris, France
5Inserm U567, Paris, France
6Université Paris 5, Faculté de Médecine René Descartes, UM 3, Paris, France
7Department of Medicine, Weill Medical College, New York, New York, USA
8Department of Biological Sciences, The Open University, Milton Keynes, UK
9Flocel Inc., Cleveland, Ohio, USA
10Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine, Cleveland, Ohio, USA

Tóm tắt

In evaluating drugs that enter or are excluded from the brain, novel pharmaceutical strategies are needed. For this reason, we have developed a humanized Dynamic In vitro Blood—Brain Barrier model (hDIV-BBB) based on a novel human brain vascular endothelial cell line (HCMEC/D3), which closely mimics the BBB in vivo. In this system, HCMEC/D3 was grown in the lumen of hollow microporous fibers and exposed to a physiological pulsatile flow. Comparison with well-established humanized DIV-BBB models (based on human brain and non-brain vascular endothelial cells co-cultured with abluminal astrocytes) demonstrated that HCMEC/D3 cells cultured under flow conditions maintain in vitro physiological permeability barrier properties of the BBB in situ even in the absence of abluminal astrocytes. Measurements of glucose metabolism demonstrated that HCMEC/D3 cells retain an aerobic metabolic pathway. Permeability to sucrose and two relevant central nervous system drugs showed that the HCMEC/D3 cells grown under dynamic conditions closely mimic the physiological permeability properties of the BBB in situ (slope = 0.93). Osmotic disruption of the BBB was also successfully achieved. Peak BBB opening in the DIV-BBB lasted from 20 to 30 mins and was completely reversible. Furthermore, the sequence of flow cessation/reperfusion in the presence of leukocytes led to BBB failure as demonstrated by a biphasic decrease in transendothelial electrical resistance. Additionally, BBB failure was paralleled by the intraluminal release of proinflammatory factors (interleukin-6 and interleukin-1β) and matrix metalloproteinase-9 (MMP-9). Pretreatment with ibuprofen (0.125 mmol/L) prevented BBB failure by decreasing the inflammatory response after flow cessation/reperfusion.

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Tài liệu tham khảo

10.1046/j.1469-7580.2002.00064.x

10.3171/jns.2003.98.4.0860

10.1111/j.1749-6632.1994.tb17359.x

10.2174/157016206777709447

10.1023/B:PHAM.0000026424.78528.11

10.1038/sj.jcbfm.9600248

10.2741/1282

Cole DJ, 1993, J Pharmacol Exp Ther, 266, 1713

10.1111/j.1528-1167.2006.00960.x

10.1016/S0006-8993(02)03167-0

Davson H, 1996, Physiology of the CSF and Blood—Brain Barriers., 49

10.1080/10623320212004

10.2741/1883

10.1007/s00441-003-0825-y

Kalaria RN, 1992, Cerebrovasc Brain Metab Rev, 4, 226

10.1007/s11064-005-8831-y

10.1016/S0006-8993(03)02689-1

10.1002/jcp.20429

10.1016/j.eplepsyres.2005.11.011

10.1002/ana.20199

10.1007/BF02968569

10.1016/S0378-5173(02)00645-2

10.1073/pnas.170282597

10.1016/j.brainres.2006.05.073

10.1016/S0006-8993(99)01872-7

10.1191/1352458503ms965oa

10.1602/neurorx.2.1.3

10.1016/S0006-8993(03)02980-9

10.1023/A:1007049806660

10.1016/j.brainres.2006.06.027

10.1007/3-211-30714-1_29

10.1007/3-211-30714-1_91

10.1016/S0006-8993(97)00829-9

10.1016/j.neuroscience.2004.06.046

10.1097/01.inf.0000131634.57368.45

10.1111/j.1528-1167.2005.00298.x

10.1111/j.1528-1157.1996.tb00586.x

10.1096/fj.04-3458fje

10.1111/j.1749-6632.1994.tb17311.x

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Thông tin xuất bản

Journal of Cerebral Blood Flow and Metabolism

Tập 28 Số 2

312-328

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