Tube-Forming Assays

Methods Mol Biol. 2016:1430:149-57. doi: 10.1007/978-1-4939-3628-1_9.

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.

Keywords: Angiogenesis; Endothelial cell; ImageJ; Matrigel; Sprouting; Tube formation.

MeSH terms

  • Cell Culture Techniques
  • Cell Movement
  • Cell Proliferation
  • Endothelial Cells / cytology
  • Endothelial Cells / ultrastructure*
  • High-Throughput Screening Assays
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Neovascularization, Physiologic*