Development, Characterization and Potential Applications of a Multicellular Spheroidal Human Blood-Brain Barrier Model Integrating Three Conditionally Immortalized Cell Lines
| Autor: | Tomomi Furihata, Yoshiyuki Yamaura, Ryo Ito, Kenta Umehara, Hidetaka Akita, Keita Kitamura, Hanae Morio, Takafumi Komori |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
THP-1 Cells Immunocytochemistry Pharmaceutical Science Inflammation In Vitro Techniques Blood–brain barrier Permeability Cell Line 03 medical and health sciences 0302 clinical medicine Spheroids Cellular medicine Humans Pharmacology Chemistry Tumor Necrosis Factor-alpha Spheroid Brain Endothelial Cells Membrane Transport Proteins Glucose analog Biological Transport General Medicine In vitro Coculture Techniques Cell biology 030104 developmental biology medicine.anatomical_structure nervous system Blood-Brain Barrier 030220 oncology & carcinogenesis Astrocytes cardiovascular system Tumor necrosis factor alpha medicine.symptom Pericytes Immortalised cell line |
| Zdroj: | Biologicalpharmaceutical bulletin. 44(7) |
| ISSN: | 1347-5215 |
| Popis: | In vitro blood-brain barrier (BBB) models are essential research tools for use in developing brain-targeted drugs and understanding the physiological and pathophysiological functions of the BBB. To develop BBB models with better functionalities, three-dimensional (3D) culture methods have gained significant attention as a promising approach. In this study, we report on the development of a human conditionally immortalized cell-based multicellular spheroidal BBB (hiMCS-BBB) model. After being seeded into non-attachment culture wells, HASTR/ci35 (astrocytes) and HBPC/ci37 cells (brain pericytes) self-assemble to form a spheroid core that is then covered with an outer monolayer of HBMEC/ci18 cells (brain microvascular endothelial cells). The results of immunocytochemistry showed the protein expression of several cellular junction and BBB-enriched transporter genes in HBMEC/ci18 cells of the spheroid model. The permeability assays showed that the hiMCS-BBB model exhibited barrier functions against the penetration of dextran (5 and 70 kDa) and rhodamine123 (a P-glycoprotein substrate) into the core. On the other hand, facilitation of 2-(N-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]amino)-2-deoxyglucose (2-NBDG; a fluorescent glucose analog) uptake was observed in the hiMCS-BBB model. Furthermore, tumor necrosis factor-alpha treatment elicited an inflammatory response in HBMEC/ci18 cells, thereby suggesting that BBB inflammation can be recapitulated in the hiMCS-BBB model. To summarize, we have developed an hiMCS-BBB model that possesses fundamental BBB properties, which can be expected to provide a useful and highly accessible experimental platform for accelerating various BBB studies. |
| Databáze: | OpenAIRE |
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