Understanding the interaction between cells and their environment is fundamental for mechanobiology. To mimic the behavior of cells in physiological and pathological conditions, synthetic substrates must have topographical and/or mechanical properties that evolve in time. Dynamic substrates mainly rely on stimuli-responsive materials where an external stimulus induces controlled variations in topography or mechanics. Herein, we describe the development of a dynamic cell culture substrate where mechanical properties are reversibly tuned in situ using magnetically-responsive superparamagnetic iron oxide nanoparticles (SPIONs).