A novel image analysis methodology for the evaluation of Angiogenesis in Matrigel assays and screening of Angiogenesis-modulating compounds

Abstract

Angiogenesis, with a prominent role in tumor growth, is a physiological process through which new blood vessels are generated from pre-existing vasculature. The effect of angiogenesis inhibition test agents to cell differentiation can be evaluated using the matrigel differentiation assay, an environment simulating in vitro the natural angiogenic process under study. Using this assay, angiogenic activity can be evaluated by the quantification of tubules, junctions and other structures of interest in the matrigel images, i.e. microphotographs of the formed cellular networks under different fields of view. In this work, we present an image analysis workflow that uses separate channels in the RGB matrigel images, each contributing different information related to angiogenic activity (tubules, junctions) and other structures (cells, lumens). The workflow is successfully showcased on the basis of a series of matrigel images generated for the evaluation of angiogenesis inhibition compounds, and results are crossvalidated with a manual expert scoring.