title: Image-based high-throughput screening for inhibitors of angiogenesis.
abstract: Automated multicolor fluorescence microscopy facilitates high-throughput quantitation of cellular parameters of complex, organotypic systems. In vitro co-cultured vascular cells form capillary-like networks that model facets of angiogenesis, making it an attractive alternative for anti-angiogenic drug discovery. We have adapted this angiogenesis assay system to a high-throughput format to enable automated image-based high-throughput screening of live primary human vascular cell co-cultures with chemical libraries for anti-angiogenic drug discovery. Protocols are described for setup of a fluorescence-based co-culture assay, live cell image acquisition, image analysis of morphological parameters, and screening data handling.
authors.affiliation: Department of Biomedicine, University of Bergen, Bergen, Norway.
authors.forename: Lasse
authors.initials: L
authors.lastname: Evensen
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Link
authors.forename: James B
authors.initials: JB
authors.lastname: Lorens
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D001562:Benzimidazoles / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D018920:Coculture Techniques; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057166:High-Throughput Screening Assays / Q000379:methods*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D032389:Myocytes, Smooth Muscle / Q000187:drug effects; D010793:Phthalazines / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000166:cytology; D011725:Pyridines / Q000494:pharmacology*; D015363:Quinolones / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013194:Staining and Labeling; D014161:Transduction, Genetic
chemical_list: C500007:4-amino-5-fluoro-3-(5-(4-methylpiperazin-1-yl)-1H-benzimidazol-2-yl)quinolin-2(1H)-one; D020533:Angiogenesis Inhibitors; D001562:Benzimidazoles; D010793:Phthalazines; D011725:Pyridines; D015363:Quinolones; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; C404768:vatalanib