title: In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling.
abstract: The blood-brain barrier (BBB) is composed of four cell populations, brain endothelial cells (BECs), pericytes, neurons, and astrocytes. Its role is to precisely regulate the microenvironment of the brain through selective substance crossing. Here we generated an in vitro model of the BBB by differentiating human induced pluripotent stem cells (hiPSCs) into all four populations. When the four hiPSC-derived populations were co-cultured, endothelial cells (ECs) were endowed with features consistent with BECs, including a high expression of nutrient transporters (CAT3, MFSD2A) and efflux transporters (ABCA1, BCRP, PGP, MRP5), and strong barrier function based on tight junctions. Neuron-derived Dll1, which activates Notch signaling in ECs, was essential for the BEC specification. We performed in vitro BBB permeability tests and assessed ten clinical drugs by nanoLC-MS/MS, finding a good correlation with the BBB permeability reported in previous cases. This technology should be useful for research on human BBB physiology, pathology, and drug development.
authors.affiliation: Laboratory of Stem Cell Differentiation, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: kohei@cira.kyoto-u.ac.jp.
authors.forename: Kohei
authors.initials: K
authors.lastname: Yamamizu
authors.affiliation: Department of Life Science Frontier, CiRA, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Mio
authors.initials: M
authors.lastname: Iwasaki
authors.affiliation: Laboratory of Stem Cell Differentiation, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Takakubo
authors.affiliation: Department of Chemistry and Biochemistry, Waseda University, Tokyo 169-8555, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Sakamoto
authors.affiliation: Laboratory of Stem Cell Differentiation, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ikuno
authors.affiliation: Laboratory of Stem Cell Differentiation, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Miyoshi
authors.affiliation: Laboratory of Stem Cell Medicine, Department of Cell Growth and Differentiation, CiRA, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Department of Chemistry and Biochemistry, Waseda University, Tokyo 169-8555, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Nakao
authors.affiliation: Department of Life Science Frontier, CiRA, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Nakagawa
authors.affiliation: Laboratory of Stem Cell Medicine, Department of Cell Growth and Differentiation, CiRA, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Haruhisa
authors.initials: H
authors.lastname: Inoue
authors.affiliation: Laboratory of Stem Cell Differentiation, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Jun K
authors.initials: JK
authors.lastname: Yamashita
mesh_terms: D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D015415:Biomarkers; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000378:metabolism*; D002199:Capillary Permeability; D002454:Cell Differentiation; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D020286:Pericytes / Q000166:cytology / Q000378:metabolism*; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D015415:Biomarkers; D051880:Receptors, Notch
keywords: Notch signaling; astrocytes; blood-brain barrier; drug kinetics; endothelial cells; induced pluripotent stem cells; neurons; pericytes; permeability; vasculature