In Vitro Assays for Endothelial Cell Functions Required for Angiogenesis: Proliferation, Motility, Tubular Differentiation, and Matrix Proteolysis

This chapter deconstructs the process of angiogenesis into its component parts in order to provide simple assays to measure discrete endothelial cell functions. The techniques described will be suitable for studying stimulators and/or inhibitors of angiogenesis and determining which aspect of the process is modulated. The assays are designed to be robust and straightforward, using human umbilical vein endothelial cells, but with an option to use other sources such as microvascular endothelial cells from various tissues or lymphatic endothelial cells. It must be appreciated that such reductionist approaches cannot cover the complexity of the angiogenic process as a whole, incorporating as it does a myriad of positive and negative signals, three-dimensional interactions with host tissues and many accessory cells including fibroblasts, macrophages, pericytes and platelets. The extent to which in vitro assays predict physiological or pathological processes in vivo (e.g., wound healing, tumor angiogenesis) or surrogate techniques such as the use of Matrigel™ plugs, sponge implants, corneal assays etc remains to be determined.