Evaluation of the immortalised mouse brain capillary endothelial cell line, b.End3, as an in vitro blood–brain barrier model for drug uptake and transport studies

Omidi, Yadollah; Campbell, Lee Ann; Barar, Jaleh; Connell, David J.; Akhtar, Saeed; Gumbleton, Mark

doi:10.1016/s0006-8993(03)03443-7

Cited by 226 publications

(205 citation statements)

References 65 publications

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“…In addition, primary cell cultures lose their differential phenotype over repeated passages. 32,33 Moreover, primary cell cultures have potential contaminations from other types of brain cells such as pericytes and astrocytes, which induce leaky barriers in the in vitro monolayer models. 8 Recently, the in vitro monolayer deriving from immortalized mouse brain endothelial cell line, bEnd3, becomes popular because of its advantages over primary cell culture, including the ability to maintain BBB characteristics over many passages, as well as easy growth and low cost.…”

Section: Introductionmentioning

confidence: 99%

Effect of Surface Charge of Immortalized Mouse Cerebral Endothelial Cell Monolayer on Transport of Charged Solutes

Yuan

2010

Ann Biomed Eng

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Abstract-Charge carried by the surface glycocalyx layer (SGL) of the cerebral endothelium has been shown to significantly modulate the permeability of the blood-brain barrier (BBB) to charged solutes in vivo. The cultured monolayer of bEnd3, an immortalized mouse cerebral endothelial cell line, is becoming a popular in vitro BBB model due to its easy growth and maintenance of many BBB characteristics over repeated passages. To test whether the SGL of bEnd3 monolayer carries similar charge as that in the intact BBB and quantify this charge, which can be characterized by the SGL thickness (L f ) and charge density (C mf ), we measured the solute permeability of bEnd3 monolayer to neutral solutes and to solutes with similar size but opposite charges: negatively charged a-lactalbumin (À11) and positively charged ribonuclease (+3). Combining the measured permeability data with a transport model across the cell monolayer, we predicted the L f and the C mf of bEnd3 monolayer, which is~160 nm and~25 mEq/L, respectively. We also investigated whether orosomucoid, a plasma glycoprotein modulating the charge of the intact BBB, alters the charge of bEnd3 monolayer. We found that 1 mg/mL orosomucoid would increase SGL charge density of bEnd3 monolayer to~2-fold of its control value.

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Section: Introductionmentioning

confidence: 99%

Effect of Surface Charge of Immortalized Mouse Cerebral Endothelial Cell Monolayer on Transport of Charged Solutes

Yuan

2010

Ann Biomed Eng

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“…Unfortunately, cerebral phenotype can be modified in these conditions. 14,23 In our procedure, collagenase/dispase digestion of isolated capillaries promoted migration of ECs. Hoechst staining and tight junction immunolabellings showed that MBCECs formed, on matrigel-coated inserts, a complete Mouse syngenic in vitro BBB model C Coisne et al monolayer of small, tightly packed, nonoverlapping and contact-inhibited cells, as expected for a vessel wall endothelium.…”

Section: Discussionmentioning

confidence: 99%

Mouse syngenic in vitro blood–brain barrier model: a new tool to examine inflammatory events in cerebral endothelium

Coisne

Dehouck

Faveeuw

et al. 2005

Laboratory Investigation

168

173

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Although cerebral endothelium disturbance is commonly observed in central nervous system (CNS) inflammatory pathologies, neither the cause of this phenomenon nor the effective participation of bloodbrain barrier (BBB) in such diseases are well established. Observations were mostly made in vivo using mouse models of chronic inflammation. This paper presents a new mouse in vitro model suitable for the study of underlying mechanistic events touching BBB functions during CNS inflammatory disturbances. This model consists of a coculture with both primary cell types isolated from mice. Mouse brain capillary endothelial cell (MBCEC)s coming from brain capillaries are in culture with their in vivo partners and form differentiated monolayers that retain endothelial markers and numerous phenotypic properties of in vivo cerebral endothelium, such as: (1) Keywords: BBB; in vitro model; coculture; tight junction; P-glycoprotein; cell adhesion moleculeThe maintenance of brain interstitial fluid homeostasis is established by the presence of the bloodbrain barrier (BBB) which can be considered as the main interface between blood and brain parenchymal cells. This endothelium can be distinguished from the other vascular beds by the presence of continuous tight junctions and the absence of fenestration or channel. Both of these characteristics reduce the unspecific transport of molecules across the BBB. The blood/brain exchanges involve specific carrier-mediated transport systems that facilitate the uptake of nutrients. 1,2 BBB damage commonly associated to inflammatory events has been reported in several central nervous system (CNS) infections and neurodegenerative diseases. [3][4][5] Although BBB permeability disturbances seem to be critical steps as they lead to brain damage, neither the cause of these phenomena nor the effective participation of BBB in such diseases are established.The cerebral vascular wall was formally considered as a barrier that isolates brain from immunecontrol. However, the presence of immune cells in the brain parenchyma in healthy animals, and their increased cerebral residence in pathogenic conditions, demonstrate that the recruitment of inflammatory cells into the CNS, through the BBB, takes place. 6,7 In fact, the presence of cell adhesion molecules (CAMs) on the BBB suggests a direct and selective interaction between blood cells and the cerebral endothelium. These molecules, first described in peripheral capillaries, may mediate leukocyte transmigration across the BBB in a multistep process. 8,9 BBB damages and leukocyte infiltration in brain parenchyma were mostly examined using in vivo mouse models of chronic inflammation, which are

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“…We used a cell culture model of BBB consisting of a coculture of mouse brain capillary endothelial cells (Omidi et al, 2003) and rat astrocytes in vitro (Dehouck et al, 1990;Deplanque et al, 2003) to assess the membrane permeability of GSNO. The permeability (25.2% of total radioactivity, at 60 mins) shows GSNO as a weak permeant molecule, indicating that GSNO may be crossing the BBB at a slow rate.…”

Section: Gsno Crosses the Endothelial Cells Slowlymentioning

confidence: 99%

S-Nitrosoglutathione Reduces Inflammation and Protects Brain against Focal Cerebral Ischemia in a Rat Model of Experimental Stroke

Khan

Sekhon

Giri

et al. 2005

J Cereb Blood Flow Metab

150

179

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Preservation of endothelial functions with low-dose nitric oxide (NO) and inhibition of excessive production of NO from inducible NO synthase (iNOS) is a potential therapeutic approach for acute stroke. Based on this hypothesis, an NO modulator, S-nitrosoglutathione (GSNO) was used, which provided neuroprotection in a rat model of focal cerebral ischemia. Administration of GSNO after the onset of ischemia reduced infarction and improved cerebral blood flow. To understand the mechanism of protection, the involvement of inflammation in ischemic brain injury was examined. Treatment with GSNO reduced the expression of tumor necrosis factor-a, interleukin-1b, and iNOS; inhibited the activation of microglia/macrophage (ED1, CD11-b); and downregulated the expression of leukocyte function-associated antigen-1 and intercellular adhesion molecule-1 in the ischemic brain. The number of apoptotic cells (including neurons) and the activity of caspase-3 were also decreased after GSNO treatment. Further, the antiinflammatory effect of GSNO on expression of iNOS and activation of NF-jB machinery in rat primary astrocytes and in the murine microglial cell line BV2 was tested. Cytokine-mediated expression of iNOS and activation of NF-jB were inhibited by GSNO treatment. That GSNO protects the brain against ischemia/reperfusion injury by modulating NO systems, resulting in a reduction in inflammation and neuronal cell death was documented by the results.

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Evaluation of the immortalised mouse brain capillary endothelial cell line, b.End3, as an in vitro blood–brain barrier model for drug uptake and transport studies

Cited by 226 publications

References 65 publications

Effect of Surface Charge of Immortalized Mouse Cerebral Endothelial Cell Monolayer on Transport of Charged Solutes

Effect of Surface Charge of Immortalized Mouse Cerebral Endothelial Cell Monolayer on Transport of Charged Solutes

Mouse syngenic in vitro blood–brain barrier model: a new tool to examine inflammatory events in cerebral endothelium

S-Nitrosoglutathione Reduces Inflammation and Protects Brain against Focal Cerebral Ischemia in a Rat Model of Experimental Stroke

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