Construction and Functional Evaluation of a Three-Dimensional Blood–Brain Barrier Model Equipped With Human Induced Pluripotent Stem Cell-Derived Brain Microvascular Endothelial Cells

• DOI: 10.1007/s11095-022-03249-3
• Publication Date: 2022-04-11T20:02:46Z
• Authors: Toshiki Kurosawa; Daiki Sako; Yuma Tega; Yasuyuki Debori; Yumi Tomihara; Kazunobu Aoyama; Yoshiyuki Kubo; Nobuyuki Amano; Yoshiharu Deguchi
• Keywords: Paracellular transport; Lucifer yellow; Claudin; Occludin; Barrier function

The purpose of this study was to construct and validate an in vitro three-dimensional blood-brain barrier (3DBBB) model system equipped with brain microvascular endothelial cells derived from human induced pluripotent stem (hiPS-BMECs).The 3D-BBB constructed by seeding hiPS-BMECs onto the capillary lane a MIMETAS OrganoPlate® 3-lane coated fibronectin/collagen IV. were incubated under continuous switchback flow OrganoFlow® for 2 days. 3D structure expression tight-junction proteins transporters confirmed immunocytochemistry. mRNA environment determined using qRT-PCR, permeability endogenous substances drugs evaluated various conditions.The proteins, including claudin-5 ZO-1, immunohistochemistry. rate constant lucifer yellow through undetectably low, indicating that paracellular transport is highly restricted tight junctions system. levels receptors differed those 2D-culture 0.2- 5.8-fold. 3D-cultured showed asymmetric substrates BCRP, CAT1 LAT1 between luminal (blood) abluminal (brain) sides. Proton-coupled symport function MCT1 also confirmed.The mimics several important characteristics BBB, expected be useful high-throughput evaluation tool development CNS drugs.