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Measurement of Angiogenic Phenotype by Use of a Two Dimensional Mesenteric Angiogenesis Assay

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Angiogenesis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 467))

Abstract

Successful therapeutic angiogenesis requires an understanding of how the milieu of growth factors available combine to form a mature vascular bed. This requires a model in which multiple physiological and cell biological parameters can be identified. The adenoviral-mediated mesenteric angiogenesis assay as described here is ideal for that purpose. Adenoviruses expressing growth factors (vascular endothelial growth factor [VEGF] and angiopoietin 1 [Ang-1]) were injected into the mesenteric fat pad of adult male Wistar rats. The clear, thin, and relatively avascular mesenteric panel was used to measure increased vessel perfusion by intravital microscopy. In addition, high-powered microvessel analysis was carried out by immunostaining of features essential for the study of angiogenesis (endothelium, pericyte, smooth muscle cell area, and proliferation), allowing functional data to be obtained in conjunction with high-power microvessel ultrastructural analysis. A combination of individual growth factors resulted in a distinct vascular phenotype from either factor alone, with all treatments increasing the functional vessel area. VEGF produced shorter, narrow, highly branched, and sprouting vessels with normal pericyte coverage. Ang-1 induced broader, longer neovessels with no apparent increase in branching or sprouting. However, Ang-1-induced blood vessels displayed a significantly higher pericyte ensheathment. Combined treatment resulted in higher perfusion, larger and less-branched vessels, with normal pericyte coverage, suggesting them to be more mature. This model can be used to show that Ang-1 and VEGF use different physiological mechanisms to enhance vascularisation of relatively avascular tissue.

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Authors and Affiliations

  1. Division of Vascular Oncology and Metastasis, DKFZ, Heidelberg, Germany

    Andrew V. Benest BSC

  2. Microvascular Research Laboratories, Bristol Heart Institute, Department of Physiology, University of Bristol, Bristol, Germany

    David O. Bates PHD

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  1. Andrew V. Benest BSC
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  2. David O. Bates PHD
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    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

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    Benest, A.V., Bates, D.O. (2009). Measurement of Angiogenic Phenotype by Use of a Two Dimensional Mesenteric Angiogenesis Assay. In: Murray, C., Martin, S. (eds) Angiogenesis Protocols. Methods in Molecular Biology, vol 467. Humana Press. https://doi.org/10.1007/978-1-59745-241-0_15

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    • DOI: https://doi.org/10.1007/978-1-59745-241-0_15

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