Establishment of a Human Blood-Brain Barrier Co-culture Model Mimicking the Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells.
In vitro models of the human blood-brain barrier (BBB) are highly desirable for drug development. This study aims to analyze a set of ten different BBB culture models based on primary cells, human induced pluripotent stem cells (hiPSCs), and multipotent fetal neural stem cells (fNSCs). We systematically investigated the impact of astrocytes, pericytes, and NSCs on hiPSC-derived BBB endothelial cell function and gene expression. The quadruple culture models, based on these four cell types, achieved BBB characteristics including transendothelial electrical resistance (TEER) up to 2,500 Ω cm2and distinct upregulation of typical BBB genes. A complex in vivo-like tight junction (TJ) network was detected by freeze-fracture and transmission electron microscopy. Treatment with claudin-specific TJ modulators caused TEER decrease, confirming the relevant role of claudin subtypes for paracellular tightness. Drug permeability tests with reference substances were performed and confirmed the suitability of the models for drug transport studies.
University Hospital Würzburg, Chair Tissue Engineering and Regenerative Medicine, 97070 Würzburg, Germany; Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases", Branch of Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 97070 Würzburg, Germany.
Antje
A
Appelt-Menzel
Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases", Branch of Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 97070 Würzburg, Germany.
Alevtina
A
Cubukova
Julius-Maximilians-University Würzburg, Institute of Anatomy and Cell Biology, Stem Cell and Regenerative Medicine Group, 97070 Würzburg, Germany.
Katharina
K
Günther
Julius-Maximilians-University Würzburg, Institute of Anatomy and Cell Biology, Stem Cell and Regenerative Medicine Group, 97070 Würzburg, Germany; Leopold-Franzens-University Innsbruck, Institute of Molecular Biology & CMBI, Department Genomics, Stem Cell Biology & Regenerative Medicine, 6020 Innsbruck, Austria.
Frank
F
Edenhofer
Charité Universitätsmedizin Berlin, Clinical Physiology & Nutritional Medicine, Department of Gastroenterology, Rheumatology & Infectious Diseases, 12203 Berlin, Germany.
Jörg
J
Piontek
Leibniz Institut für Molekulare Pharmakologie, 13125 Berlin, Germany.
Gerd
G
Krause
University Hospital Würzburg, Women's Hospital and Polyclinic, 97080 Würzburg, Germany.
Tanja
T
Stüber
University Hospital Würzburg, Chair Tissue Engineering and Regenerative Medicine, 97070 Würzburg, Germany; Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases", Branch of Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 97070 Würzburg, Germany.
Heike
H
Walles
AIT Austrian Institute of Technology GmbH, Competence Center Health and Bioresources, Competence Unit Molecular Diagnostics, 1190 Vienna, Austria.
Winfried
W
Neuhaus
University Hospital Würzburg, Chair Tissue Engineering and Regenerative Medicine, 97070 Würzburg, Germany; Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases", Branch of Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, 97070 Würzburg, Germany. Electronic address: marco.metzger@igb.fraunhofer.de.
Marco
M
Metzger
D001812:Blood-Brain Barrier / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D002199:Capillary Permeability; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D005333:Fetus / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D000069336:Neurovascular Coupling; D010599:Pharmacokinetics; D019108:Tight Junctions / Q000378:metabolism / Q000648:ultrastructure
blood-brain barrier (BBB) model; human induced pluripotent stem cells (hiPSCs); multipotent fetal neural stem cells (fNSCs); neurovascular unit in vitro