Development, Characterization and Potential Applications of a Multicellular Spheroidal Human Blood–Brain Barrier Model Integrating Three Conditionally Immortalized Cell Lines

• Kitamura Keita

  Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Chiba University Laboratory of Clinical Pharmacy & Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences

• Umehara Kenta

  Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Chiba University

• Ito Ryo

  Research Center of Neurology, Ono Pharmaceutical Co., Ltd.

• Yamaura Yoshiyuki

  Pharmacokinetic Research Laboratories, Ono Pharmaceutical Co., Ltd.

• Komori Takafumi

  Drug Metabolism and Pharmacokinetics Tsukuba, Tsukuba Research Laboratories, Eisai Co., Ltd.

• Morio Hanae

  Laboratory of Clinical Pharmacy & Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences

• Akita Hidetaka

  Laboratory of DDS Design and Drug Disposition, Graduate School of Pharmaceutical Sciences, Chiba University

• Furihata Tomomi

  Laboratory of Clinical Pharmacy & Experimental Therapeutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences

Abstract

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.

Journal

• Biological and Pharmaceutical Bulletin

  Biological and Pharmaceutical Bulletin 44 (7), 984-991, 2021-07-01

  The Pharmaceutical Society of Japan

Keywords

• blood–brain barrier
• <i>in vitro</i> model
• immortalized cell
• spheroid
• drug development
• central nervous system disease

Details

• CRID: 1390851497214023808
• NII Article ID: 130008060355
• NII Book ID: AA10885497
• DOI: 10.1248/bpb.b21-00218
• ISSN: 13475215
• ISSN: 09186158
• NDL BIB ID: 031534979
• PubMed: 33896887
• Web Site: http://id.ndl.go.jp/bib/031534979
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I031534979
• Web Site: https://www.jstage.jst.go.jp/article/bpb/advpub/0/advpub_b21-00218/_pdf
• Web Site: https://search.jamas.or.jp/link/ui/2022141300
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed