Investigating In Vitro Angiogenesis by Computer-Assisted Image Analysis and Computational SimulationVascular Morphogenesis

In vitro assays that stimulate the formation of capillary-like structures by EC have become increasingly popular, because they allow the study of the EC's intrinsic ability to self-organize to form vascular-like patterns. Here we describe a widely applied protocol involving the use of basement membrane matrix (Matrigel) as a suitable environment to induce an angiogenic phenotype in cultured EC. EC differentiation on basement membrane matrix is a highly specific process, which recapitulates many steps in blood vessel formation and for this reason it is presently considered as a reliable in vitro tool to identify factors with potential antiangiogenic or pro-angiogenic properties. The morphological features of the obtained cell patterns can also be accurately quantified by computer-assisted image analysis and the main steps of such a procedure will be here outlined and discussed. The dynamics of in vitro EC self-organization is a complex biological process, involving a network of interactions between a high number of cells. For this reason, the combined use of in vitro experiments and computational modeling can represent a key approach to unravel how mechanical and chemical signaling by EC coordinates their organization into capillary-like tubes. Thus, a particularly helpful approach to modeling is also briefly described together with examples of its application.