Use of the mouse aortic ring assay to study angiogenesis

Marianne Baker, Stephen Robinson, Tanguy Lechertier, Paul R Barber, Bernardo Tavora, Gabriela D'Amico, Dylan T Jones, Boris Vojnovic, Kairbaan Hodivala-Dilke

Research output: Contribution to journalArticlepeer-review

311 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)89-104
Number of pages16
JournalNature Protocols
Volume7
Issue number1
DOIs
Publication statusPublished - Jan 2012

Keywords

  • Microscopy, Fluorescence
  • Cell Movement
  • Animals
  • Microscopy, Phase-Contrast
  • Neovascularization, Physiologic
  • Aorta
  • Tissue Culture Techniques
  • Mice, Inbred C57BL
  • Mice
  • Cell Proliferation

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