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. Oour protocol can be applied to aortic rings from embryonic stage Ee18 through to adulthood and can incorporate genetic manipulation, treatment with growth factors, drugs or siRNArna. 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. assay takes 6-14 d to complete, depending on the age of the mice, treatments applied and whether immunostaining is performed. © 2012 Nature America, Inc. All rights reserved.