Geometric confinement influences cellular mechanical properties II -- intracellular variances in polarized cells

During migration, asymmetrically polarized cells achieve motion by coordinating the protrusion and retraction of their leading and trailing edges, respectively. Although it is well known that local changes in the dynamics of actin cytoskeleton remodeling drive these processes, neither the cytoskeletal rheological properties of these migrating cells are well quantified nor is it understand how these rheological properties are regulated by underlying molecular processes. In this report, we have used soft lithography to create morphologically polarized cells in order to examine rheological differences between the front and rear zone of an NIH 3T3 cell posed for migration. In addition, we trapped superparamagnetic beads with optical tweezers and precisely placed them at specific locations on the immobilized cells. The beads were then allowed to endocytose overnight before magnetic tweezers experiments were performed to measure the local rheological properties of the leading and trailing edges. Our results indicate that the leading edge has an approximately 1.9 times higher shear modulus than the trailing edge and that this increase in shear modulus correlates with a greater density of filamentous actin, as measured by phalloidin-staining observed through quantitative 3D microscopy.