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The in vitro methods to recapitulate environmental cues around cells are usually optimized to test a specific property of the environment (biochemical nature or the stiffness of the extracellular matrix (ECM), or nanotopography) for its capability to induce defined cell behaviors (lineage commitment, migration). Approaches that combine different environmental cues in 3D to assess the biological response of cells to the spatial organization of different biophysical and biochemical cues are growingly being developed. It is demonstrated how the lamination of through‐hole polymeric biofunctionalized membranes can be implemented to create complex bona fide microniches with differential 3D environmental properties using photoactive materials. The approach enables to create microniches ranging in size from single cells to cell aggregates. They are biofunctionalized in 3D simultaneously with topographical featured …