title: Identification of neuronal and angiogenic growth factors in an in vitro blood-brain barrier model system: Relevance in barrier integrity and tight junction formation and complexity.
abstract: We previously demonstrated that the co-cultivation of endothelial cells with neural cells resulted in an improved integrity of the in vitro blood-brain barrier (BBB), and that this model could be useful to evaluate the transport properties of potential central nervous system disease drugs through the microvascular brain endothelial. In this study we have used real-time PCR, fluorescent microscopy, protein arrays and enzyme-linked immunosorbent assays to determine which neural- and endothelial cell-derived factors are produced in the co-culture and improve the integrity of the BBB. In addition, a further improvement of the BBB integrity was achieved by adjusting serum concentrations and growth factors or by the addition of brain pericytes. Under specific conditions expression of angiogenic, angiostatic and neurotrophic factors such as endostatin, pigment epithelium derived factor (PEDF/serpins-F1), tissue inhibitor of metalloproteinases (TIMP-1), and vascular endothelial cell growth factor (VEGF) closely mimicked the in vivo situation. Freeze-fracture analysis of these cultures demonstrated the quality and organization of the endothelial tight junction structures and their association to the two different lipidic leaflets of the membrane. Finally, a multi-cell culture model of the BBB with a transendothelial electrical resistance up to 371 (±15) Ω×cm2was developed, which may be useful for preliminary screening of drug transport across the BBB and to evaluate cellular crosstalk of cells involved in the neurovascular unit.
authors.affiliation: REPAIR-lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz and European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Langenbeckstr. 1, 55131 Mainz, Germany. Electronic address: freesec@uni-mainz.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Freese
authors.affiliation: REPAIR-lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz and European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Langenbeckstr. 1, 55131 Mainz, Germany. Electronic address: hanada346@gmail.com.
authors.forename: Sanshiro
authors.initials: S
authors.lastname: Hanada
authors.affiliation: Institute of Pathology and Neuropathology, University of Tuebingen, Geschwister-Scholl-Platz, 72074 Tuebingen, Germany. Electronic address: Petra.Fallier-Becker@med.uni-tuebingen.de.
authors.forename: Petra
authors.initials: P
authors.lastname: Fallier-Becker
authors.affiliation: REPAIR-lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz and European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Langenbeckstr. 1, 55131 Mainz, Germany. Electronic address: kirkpatrick@uni-mainz.de.
authors.forename: C James
authors.initials: CJ
authors.lastname: Kirkpatrick
authors.affiliation: REPAIR-lab, Institute of Pathology, University Medical Center of the Johannes Gutenberg University Mainz and European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Langenbeckstr. 1, 55131 Mainz, Germany. Electronic address: runger@uni-mainz.de.
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Unger
mesh_terms: D042501:Angiogenic Proteins / Q000378:metabolism*; D000818:Animals; D001812:Blood-Brain Barrier / Q000166:cytology / Q000378:metabolism*; D002199:Capillary Permeability; D002450:Cell Communication; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D017097:Electric Impedance; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D009414:Nerve Growth Factors / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D000069336:Neurovascular Coupling; D010641:Phenotype; D015398:Signal Transduction; D034421:Sus scrofa; D062725:Tight Junction Proteins / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism*
chemical_list: D042501:Angiogenic Proteins; D009414:Nerve Growth Factors; D062725:Tight Junction Proteins
keywords: Brain pericytes; Endothelial-neuron crosstalk; Neurovascular unit; Pentraxin-3; Porcine brain microvascular endothelial cells