In Vitro Tube Formation Assays in Matrigel

Vasculature development is a combination of complex processes that require precise coordination of multiple cell types, through time and space, to generate functional blood-carrying vessels. Moreover, vasculature development can be altered when normal physiological conditions are disrupted, such as in cancer, and means to study blood vessels are of great importance. While the gold standard to explore these processes is the use of in vivo animal models, they are costly and time-consuming, and it is often difficult to dissect the molecular mechanisms involved. Thus, there are several ways to deconstruct vasculature development in vitro, in order to produce tunable systems that lead to a better understanding of cellular and molecular communication between different cell types involved, such as endothelial cells and supporting mesenchymal cells. In this method chapter, we will go into detail for one of the most popular ways of studying vasculature development in the context of cancer, which is the application of Matrigel to study tube formation of various cell types involved with vasculature development. We will provide step-by-step instructions to perform mono- and co-cultures of the major cells involved with the production of vasculature, how the results of these assays can be interpreted, and some advice to avoid common mistakes associated with Matrigel tube formation assays.