The aim of the pipeline is to extract and analyze relevant information from tube formation assay images (obtained from fluorescent microscopy).
One of the most widely used in vitro assays to model the reorganization stage of angiogenesis is the tube formation assay. The assay measures the ability of endothelial cells, plated at subconfluent densities with the appropriate extracellular matrix support, to form capillary-like structures (a.k.a tubes). Scientists typically employ this assay to determine the ability of various compounds to promote or inhibit tube formation. Compounds that are able to inhibit tube formation could be useful in various diseases, such as cancer, where tumors stimulate new blood vessel formation to receive oxygen and nutrients in order to grow beyond a relatively small size. Upon plating, endothelial cells attach and generate mechanical forces on the surrounding extracellular support matrix to create tracks or guidance pathways that facilitate cellular migration. The resulting cords of cells will eventually form hollow lumens.
Ongoing research is focused on how specific molecules in the “matrix-integrin-cytoskeletal signaling axis” are involved in the eventual assembly into three-dimensional vascular networks. Once formed, these interconnected networks are usually maintained for approximately 24 hours. Tube formation is typically quantified by measuring the number, length, or area of these capillary-like structures in two-dimensional microscope images of the culture dish.