title: Endothelial cell tube formation assay for the in vitro study of angiogenesis.
abstract: Angiogenesis is a vital process for normal tissue development and wound healing, but is also associated with a variety of pathological conditions. Using this protocol, angiogenesis may be measured in vitro in a fast, quantifiable manner. Primary or immortalized endothelial cells are mixed with conditioned media and plated on basement membrane matrix. The endothelial cells form capillary like structures in response to angiogenic signals found in conditioned media. The tube formation occurs quickly with endothelial cells beginning to align themselves within 1 hr and lumen-containing tubules beginning to appear within 2 hr. Tubes can be visualized using a phase contrast inverted microscope, or the cells can be treated with calcein AM prior to the assay and tubes visualized through fluorescence or confocal microscopy. The number of branch sites/nodes, loops/meshes, or number or length of tubes formed can be easily quantified as a measure of in vitro angiogenesis. In summary, this assay can be used to identify genes and pathways that are involved in the promotion or inhibition of angiogenesis in a rapid, reproducible, and quantitative manner.
authors.affiliation: Department of Biology, American University; decicco@american.edu.
authors.forename: Katie L
authors.initials: KL
authors.lastname: DeCicco-Skinner
authors.affiliation: Department of Biology, American University.
authors.forename: Gervaise H
authors.initials: GH
authors.lastname: Henry
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, NIH.
authors.forename: Christophe
authors.initials: C
authors.lastname: Cataisson
authors.affiliation: Department of Biology, American University.
authors.forename: Tracy
authors.initials: T
authors.lastname: Tabib
authors.affiliation: Department of Biology, American University.
authors.forename: J Curtis
authors.initials: JC
authors.lastname: Gwilliam
authors.affiliation: Department of Biology, American University.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Watson
authors.affiliation: Department of Biology, American University.
authors.forename: Erica M
authors.initials: EM
authors.lastname: Bullwinkle
authors.affiliation: Department of Biology, American University.
authors.forename: Lauren
authors.initials: L
authors.lastname: Falkenburg
authors.affiliation: Department of Biology, American University.
authors.forename: Rebecca C
authors.initials: RC
authors.lastname: O'Neill
authors.affiliation: Department of Biology, American University.
authors.forename: Adam
authors.initials: A
authors.lastname: Morin
authors.affiliation: Laboratory of Cancer Biology and Genetics, National Cancer Institute, NIH.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Wiest
mesh_terms: D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D017077:Culture Media, Conditioned; D042783:Endothelial Cells / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D018919:Neovascularization, Physiologic / Q000502:physiology*
chemical_list: D017077:Culture Media, Conditioned