Shape memory polymer (SMP) substrates compatible with cell culture are presented that undergo programmed changes in surface topography on command to direct cell behavior. SMPs are a class of active materials that can be programmed to transition from a stable temporary shape to a permanent shape by a triggering mechanism, such as heat. Here, an SMP surface topography was programmed to transition from parallel micron-scale grooves to a flat surface. C3H/10T1/2 cells were allowed to adhere and spread on the temporary topography at 30°C. Topographic transition was then triggered by transfer to 37°C. Changes in substrate topography were found to direct cell behavior, as assessed by analysis of cell alignment, while high cell viability was maintained. This first example of active cell culture substrates should have great potential in the design of unique approaches for advanced investigation of mechanotransduction and cell biomechanical function and softmatter physics.