title: Use of the mouse aortic ring assay to study angiogenesis.
abstract: Here we provide a protocol for quantitative three-dimensional ex vivo mouse aortic ring angiogenesis assays, in which developing microvessels undergo many key features of angiogenesis over a timescale similar to that observed in vivo. The aortic ring assay allows analysis of cellular proliferation, migration, tube formation, microvessel branching, perivascular recruitment and remodeling-all without the need for cellular dissociation-thus providing a more complete picture of angiogenic processes compared with traditional cell-based assays. Our protocol can be applied to aortic rings from embryonic stage E18 through to adulthood and can incorporate genetic manipulation, treatment with growth factors, drugs or siRNA. This robust assay allows assessment of the salient steps in angiogenesis and quantification of the developing microvessels, and it can be used to identify new modulators of angiogenesis. The assay takes 6-14 d to complete, depending on the age of the mice, treatments applied and whether immunostaining is performed.
authors.affiliation: Adhesion and Angiogenesis Laboratory, Centre for Tumour Biology, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London, UK. m.baker@qmul.ac.uk
authors.forename: Marianne
authors.initials: M
authors.lastname: Baker
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Robinson
authors.forename: Tanguy
authors.initials: T
authors.lastname: Lechertier
authors.forename: Paul R
authors.initials: PR
authors.lastname: Barber
authors.forename: Bernardo
authors.initials: B
authors.lastname: Tavora
authors.forename: Gabriela
authors.initials: G
authors.lastname: D'Amico
authors.forename: Dylan T
authors.initials: DT
authors.lastname: Jones
authors.forename: Boris
authors.initials: B
authors.lastname: Vojnovic
authors.forename: Kairbaan
authors.initials: K
authors.lastname: Hodivala-Dilke
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000502:physiology*; D002465:Cell Movement; D049109:Cell Proliferation; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D018919:Neovascularization, Physiologic / Q000502:physiology*; D046509:Tissue Culture Techniques