Self-standing nanocomposite films based on biopolymers and functional nanostructures have been widely used due to their potential applications as active elements in biomedical devices. The coupling between chitosan (CHI) and alginate (ALG) multilayered films and magnetic nanoparticles (MNPs) allowed to fabricate magnetic responsive freestanding membranes with a high structural control along the thickness, using the layer-by-layer (LbL) methodology. The mechanical characterization evidenced a trend for an increase of both Young modulus, and ultimate tensile strength with the inclusion of MNPs, or by cross-linking with genipin. Additionally, the multilayered membranes exhibited shape memory properties triggered by hydration. The in vitro biological performance studies showed that cells were more viable and adherent with higher proliferation rates when MNPs were included in the membranes. Our results suggested the potential of the developed magneto-active freestanding membranes for biomedical applications, such as in tissue engineering and biomedical applications.