A microfabricated platform probing cytoskeleton dynamics using multidirectional topographical cues

Abstract

Cell migration, which involves complicated coordination of cytoskeleton elements and regulatory molecules, plays a central role in a large variety of biological processes from development, immune response to tissue regeneration. However, conventional methods to study in vitro cell migration are often limited to stimulating a cell along a single direction or at a single location. This restriction prevents a deeper understanding of the fundamental mechanisms that control the spatio-temporal dynamics of cytoskeleton. Here we report a novel microfabricated platform that enables a multi-directional stimulation to a cell using topographical cues. In this device, cells were seeded on a grid-patterned topographically structured surface composed of 2 μm wide and 2 μm high straight ridges. Because the size of a unit grid was smaller than a single cell, each cell was simultaneously experiencing contact guidance leading to different directions. The device showed that healthy cells preferred to align and migrate in the direction of the longer side of the grid. But cells with impaired intracelluar tension force generation exhibited multiple uncoordinated cell protrusions along guiding ridges in all directions. Our results demonstrate the importance of actomyosin network in long-range communication and regulation of local actin polymerization activities. This platform will find wide applications in investigations of signal transduction and regulation process in cell migration.