Heterostructures and interfacial defects in a 40-nm-thick SrTiO3 (STO) film grown epitaxially on a single-crystal MgO (001) were investigated using aberration-corrected scanning transmission electron microscopy and geometric phase analysis. The interface of STO/MgO was found to be of the typical domain-matching epitaxy with a misfit dislocation network having a Burgers vector of ½ aSTO ⟨100⟩. Our studies also revealed that the misfit dislocation cores at the heterogeneous interface display various local cation arrangements in terms of the combination of the extra-half inserting plane and the initial film plane. The type of the inserting plane, either the SrO or the TiO2 plane, alters with actual interfacial conditions. Contrary to previous theoretical calculations, the starting film planes were found to be dominated by the SrO layer, i.e., a SrO/MgO interface. In certain regions, the starting film planes change to the TiO2/MgO interface because of atomic steps at the MgO substrate surface. In particular, four basic misfit dislocation core configurations of the STO/MgO system have been identified and discussed in relation to the substrate surface terraces and possible interdiffusion. The interface structure of the system in reverse—MgO/STO—is also studied and presented for comparison.