Abstract
Angiogenesis involves the generation of new blood vessels from the existing vasculature and is dependent on many growth factors and signaling events. In vivo angiogenesis is dynamic and complex, meaning assays are commonly utilized to explore specific targets for research into this area. Tube-forming assays offer an excellent overview of the molecular processes in angiogenesis. The Matrigel tube forming assay is a simple-to-implement but powerful tool for identifying biomolecules involved in angiogenesis. A detailed experimental protocol on the implementation of the assay is described in conjunction with an in-depth review of methods that can be applied to the analysis of the tube formation. In addition, an ImageJ plug-in is presented which allows automatic quantification of tube images reducing analysis times while removing user bias and subjectivity.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Adams RH, Alitalo K (2007) Molecular regulation of angiogenesis and lymphangiogenesis. Nat Rev Mol Cell Biol 8:464–478
Heath VL, Bicknell R (2009) Anticancer strategies involving the vasculature. Nat Rev Clin Oncol 6:395–404
Auerbach R, Lewis R, Shinners B et al (2003) Angiogenesis assays: a critical overview. Clin Chem 49:32–40
Bikfalvi A, Cramer EM, Tenza D et al (1991) Phenotypic modulations of human umbilical vein endothelial cells and human dermal fibroblasts using two angiogenic assays. Biol Cell 72:275–278
Connolly JO, Simpson N, Hewlett L et al (2002) Rac regulates endothelial morphogenesis and capillary assembly. Mol Cell Biol 13:2474–2485
Kaur S (2011) Characterisation of the expression profile and endothelial function of Rho GTPase RhoJ. PhD Thesis. University of Birmingham
Donovan D, Brown NJ, Bishop ET et al (2001) Comparison of three in vitro human “angiogenesis” assays with capillaries formed in vivo. Angiogenesis 4:113–121
Staton CA, Reed MWR, Brown NJ (2009) A critical analysis of current in vitro and in vivo angiogenesis assays. Int J Exp Pathol 90:195–221
Nguyen DHT, Stapleton SC, Yang MT et al (2013) Biomimetic model to reconstitute angiogenic sprouting morphogenesis in vitro. Proc Natl Acad Sci U S A 110:6712–6717
Kang H, Bayless KJ, Kaunas R (2008) Fluid shear stress modulates endothelial cell invasion into three-dimensional collagen matrices. Am J Physiol Heart Circ Physiol 3120:2087–2097
Khoo CP, Micklem K, Watt SM (2011) A comparison of methods for quantifying angiogenesis in the Matrigel assay in vitro. Tissue Eng Part C Methods 17:895–906
Landini G (2008) Advanced shape analysis with ImageJ. In: Second ImageJ user developer conference proceedings. Luxembourg. p 116–121
Carpentier G, Martinelli M, Courty J et al (2012) Angiogenesis analyzer for ImageJ. In: Fourth ImageJ user developer conference proceedings. Mondorf-les-Bains, Luxembourg, p 198–201
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Brown, R.M., Meah, C.J., Heath, V.L., Styles, I.B., Bicknell, R. (2016). Tube-Forming Assays. In: Martin, S., Hewett, P. (eds) Angiogenesis Protocols. Methods in Molecular Biology, vol 1430. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3628-1_9
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3628-1_9
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3626-7
Online ISBN: 978-1-4939-3628-1
eBook Packages: Springer Protocols