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The new in vitro blood-brain barrier (BBB) model under shear forces with multi-culture of BBB components in multiple sclerosis
Author(s): ,
Y. Takeshita
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
,
S. Fujikawa
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
,
T. Maeda
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
,
F. Shimizu
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
,
Y. Sano
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
T. Kanda
Affiliations:
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
ECTRIMS Online Library. Takeshita Y. Oct 12, 2018; 228906; P1065
Yukio Takeshita
Yukio Takeshita
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Abstract: P1065

Type: Poster Sessions

Abstract Category: Pathology and pathogenesis of MS - Experimental models

Background: The blood-brain barrier (BBB), which is mainly composed of endothelial cells , pericytes and astrocytes, play crucial roles in the pathogenesis of inflammatory neurological disorders as multiple sclerosis (MS). It has not been adequately investigated how auto-immune leukocytes infiltrate across the BBB in multiple sclerosis (MS) because of the lack of the in vitro models that are specialized to the following four essential properties. First, cells that demonstrate human BBB properties are utilized. Second, the model incorporates multi-culture of BBB components : endothelium, astrocytes and pericytes. Third, it incorporates physiological shear stress. Fourth, it allows the transmigration of leukocytes which can be recovered for further analysis.
Aim: Our aims in this study are to generate a BBB model with these four properties, and using this model, to evaluated influence of activation of BBB components with MS-related cytokines and chemokine for inflammatory cells.
Methods: Human astrocytes were cultured on abluminal side of the membrane (3µm pores) with human pericytes on luminal side. Sheet-like clusters of human endothelial cells were transferred onto the pericytes by means of the UpCell technology. Cells were activated by TNF-α and IFN-γ for 24hr at 37°C. After CXCL12 and CCL2 were added in luminal and abluminal side respectively, normal human peripheral blood mononuclear cells flowed onto luminal side with shear force. Total migrated cells were quantified and phenotyped by flow cytometry.
Results: Activation by cytokines and chemokines increased transmigration of total migrated cells, CD4+ and CD8+ T cells, CD19+ B cells, and CD14+ monocyte.
Conclusion: We successfully generated the first multi-cultured BBB model that allows the transmigration of leukocytes under shear forces. This model will provide reproducible assays for barrier regulations with robust results, which will enable further defining the relationships between inflammatory cells and the BBB components in MS.
Disclosure: The authors have no financial conflicts of interest to disclose concerning the presentation.

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