Magnetite (Fe3O4) thin films were deposited on MgO (0 0 1), SrTiO3 (0 0 1), LaAlO3 (0 0 1) single crystal substrates as well on as silicon and amorphous glass in order to study the effect of the substrate on their magnetic properties, mainly the magnetic anisotropy. We have performed a structural, morphological and compositional characterization by X-ray diffraction, atomic force microscopy and Rutherford backscattering ion channeling in oxygen resonance mode. The magnetic anisotropy has been investigated by vectorial magneto-optical Kerr effect. The results indicate that the magnetic anisotropy is especially influenced by the substrate-induced microstructure. In-plane isotropy and uniaxial anisotropy behavior have been observed on silicon and glass substrates, respectively. The transition between both behaviors depends on grain size. For LaAlO3 substrates, in which the lattice mismatch between the Fe3O4 films and the substrate is significant, a weak in-plane fourfold magnetic anisotropy is induced. However when magnetite is deposited on MgO (0 0 1) and SrTiO3 (0 0 1) substrates, a well-defined fourfold in-plane magnetic anisotropy is observed with easy axes along [1 0 0] and [0 1 0] directions. The magnetic properties on these two latter substrates are similar in terms of magnetic anisotropy and coercive fields. (C) 2015 Elsevier B.V. All rights reserved.