Localized mechanical stimulation of single cells with engineered spatio-temporal profile

Abstract

In vivo, cells are frequently exposed to multiple mechanical stimuli arising from the extracellular microenvironment, with a deep impact on many biological functions. On the other hand, current methods for mechanobiology do not allow one to easily replicate in vitro the complex spatio-temporal profile of such mechanical signals. Here we introduce a new platform for studying the mechanical coupling between single cells and a dynamic extracellular environment, based on active substrates for cell culture made of Fe-coated polymeric micropillars. Under the action of quasi-static external magnetic fields, each group of pillars produces synchronous mechanical stimuli at different points of the cell membrane, thanks to the highly controllable pillars' deflection. This method allows one to apply complex stress fields, resulting in the parallel application of localized forces with tunable intensity and temporal profile. The platform has been validated by studying the cellular response to periodic stimuli in NIH3T3 fibroblasts. We find that low-frequency mechanical stimulation affects the actin cytoskeleton, nuclear morphology, and H2B core-histone dynamics and induces MKL transcription-cofactor translocation from nucleus to cytoplasm. The unique capability of the proposed platform to apply stimuli with a tunable temporal profile and high parallelism on a cell culture holds great potential for the investigation of mechanotransduction mechanisms in cells and tissues.