Cell response to exogenous cues is the result of a complex integration of multiple biochemical/biophysical signals, which might occur simultaneously and might be characterized by specific spatial and temporal patterns. Among these signals, surface topography plays an important role in affecting cell functions and fate. However, the current understanding of the interplay between cells and topography relies on static environment. Here the intrinsic light-responsive properties of azopolymers and the versatility of laser-based confocal microscope technique is exploited, aiming to induce spatio-temporal dynamic topographic changes in situ during cell culture. Diverse patterns can be designed on cell-populated azopolymer films with high control on time, space, and on-off signal modification. The technique proposed in this study enables the development of synthetic platforms that finely control cell orientation and migration both in time and space. The results may pave the way to unravel complex processes involved in cell-topography interactions, thus allowing to define the spatio-temporal features that most effectively influence cell functions.