Cells are able to sense and react to their physical environment by translating a mechanical cue into an intracellular biochemical signal that triggers biological and mechanical responses. This process, called mechanotransduction, controls essential cellular functions such as proliferation and migration. The cellular response to an external mechanical stimulation has been investigated with various static and dynamic systems, so far limited to global deformations or to local stimulation through discrete substrates. To apply local and dynamic mechanical constraints at the single cell scale through a continuous surface, we have developed and modelled magneto-active substrates made of magnetic micro-pillars embedded in an elastomer. Constrained and unconstrained substrates are analysed to map surface stress resulting from the magnetic actuation of the micro-pillars and the adherent cells. These substrates have a rigidity in the range of cell matrices, and the magnetic micro-pillars generate local forces in the range of cellular forces, both in traction and compression. As an application, we followed the protrusive activity of cells subjected to dynamic stimulations. Our magneto-active substrates thus represent a new tool to study mechanotransduction in single cells, and complement existing techniques by exerting a local and dynamic stimulation, traction and compression, through a continuous soft substrate.Living cells have a sense of touch, which means that they are able to feel, respond and adapt to the mechanical properties of their environment. The process by which cells convert mechanical signals into biochemical signals is called mechanotransduction. Defects in the mechanotransduction pathways are implicated in numerous diseases ranging from atherosclerosis and osteoporosis to cancer progression and developmental disorders 1,2 . Since the 1990s, different static studies focused on mechanosensing have shown that cells can migrate along the rigidity gradient direction 3 and that stem cells can differentiate in vitro according to their substrate's stiffness 4 and geometry 5 . The interplay between a mechanical force and the reinforcement of cell adhesion has also been documented 6,7 . In their natural environment, cells face a complex and dynamic mechanical environment. Cyclic strain can induce reorientation of adherent cells and affect cell growth depending on the temporal and spatial properties of the mechanical stimulation [8][9][10][11] . The relevant timescales span from the milli-second for the stretching of mechanosensitive proteins, minutes for mechanotransduction signalling to hours for global morphological changes and even longer for adapting cell functions 12 . Taken together, previous works have shown that cells are sensitive to both the spatial and temporal signatures of mechanical stimuli. In order to study mechanotransduction, it is thus essential to stimulate cells with mechanical cues controlled both spatially and temporally.To address this topic, various methods have been proposed to exert experim...