Kinesin-Driven Active Substrate Giving Stochastic Mechanical Stimuli to Cells for Characterization

We present a new platform to give stochastic mechanical stimuli to cells for their characterization. There nano- and micrometer scaled fluctuations are generated by an engineered motor protein system of kinesin-microtubules (MTs) on a solid surface. Cells have abilities to deform in many ways during homeostatic metabolism, tissue forming processes, cancer developments, and so on. Namely, cells in biological tissues are exposed to noise-like stochastic movements at nano- and micrometer-scales, which mainly come from the mechanical environment surrounding the cells. Although cells seem to have the potential to respond to such types of mechanical stimuli, the influences on cellular behaviors are poorly understood. As a first attempt to verify an effect of noise-like mechanical stimuli in vitro, we prepared a system to give stochastic mechanical stimuli to cells using a technique of in vitro motility assay for a kinesin-MT system. An active substrate was obtained by integrating movements of MTs on a kinesin-coated glass surface via cross-linkage, and stochastic mechanical stimuli at the cell-scale were successfully applied to the seeded cells. There, traveling distances of the cells over one cell length were observed until they started to adhere. When metastatic melanoma cells were exposed to the stochastic mechanical stimuli, unusually long protrusions or extensions of cell bodies were observed. Cellular aggregations were also promoted through the movements on this active substrate which could disturb the landing and enhance the collisions of the cells. This approach giving mechanical stimuli to cells in a stochastic manner at nano- and micrometer-scales might allow us to uncover unknown behaviors of cells, which might contribute to research fields requiring our understanding on the mechanical nature of cells, such as cancer diagnosis and regenerative medicine.