The role of vimentin in cell migration under confinement
Cell migration is a fundamental biological process which requires an active cytoskeleton remodelling. During migration, cells often perform long distance displacements passing through obstructive 3D tissues, constrictions, or even in between other cells and therefore continuously need to adjust their shape. The intermediate filaments are the most stretchable components of the cytoskeleton and are organized as a network comprising both cytoplasmatic and nuclear proteins. It is not well established if and how alterations in the extracellular matrix can have an impact in intermediate filaments organization and cell movement. Considering this scenario, we subjected cells to confinement using microfabricated channels with the objective to analyse cell migration and the cytoplasmatic intermediate filament vimentin network. In this given context we analysed two models of cell migration- the low adherent amoeboid performed by Jurkat cell lines and the mesenchymal migration using RPE-1 cells. Both cell lines were stable transfected with wild type vimentin coupled with GFP. Cell movement in the channels was recorded by fluorescence and phase contrast video microscopy for 24 hours under controlled humidity and temperature. The acquired data was analysed by a custom-built cell tracking software. The analysed parameters during both amoeboid and mesenchymal cell migration were the speed, persistence as well as the localization of vimentin network and cell morphology (length, shape, symmetry, nuclear position, nuclear shape).