The rigidity in fibrin gels as a contributing factor to the dynamics of in vitro vascular cord formation

While the formation of vascular cords in in vitro angiogenesis assay is commonly used to test the angiogenic properties of many molecular or cellular components, an extensive characterisation of the dynamics of this process is still lacking. Up to now, quantitative studies only focused on the resulting capillary structures characterised through static morphometric approaches. We therefore propose in this paper a rather extensive characterisation aiming to identify different stages in the dynamics of this process, through the investigation of the influence of the rigidity of the fibrin extracellular matrix on the growth of the vascular cords. Using time lapse videomicroscopy, the time evolution of relevant morphodynamical parameters has been considered both at the cell level and at the cell population level. At the cell level, a trajectography analysis of individual cells observed in different locations of the growing network has been conducted and analysed using a random walk model. From image sequence analysis and segmentation i.e. extraction of the boundaries of the lacunae formed through matrix degradation and cell tractions, the evolution of the lacunae surface has been precisely quantified, revealing different phases and transitions in the growth patterns. Our results indicate that the rigidity of the extracellular fibrin matrix strongly influences the different stages, i.e. the dynamics of the angiogenic process. Consequently, optimal rigidity conditions for the formation of stable vascular cord networks could be identified in the context of our experiments.