It is well known that the use of continuous reinforcing fibers can largely improve the typical low in-plane mechanical properties of 3D-printed parts. However, there is very limited research on the characterization of the interlaminar fracture toughness of 3D-printed composites. In this study, we investigated the feasibility of determining the mode I interlaminar fracture toughness of 3D-printed cFRP composites with multidirectional interfaces. First, elastic calculations and different FE simulations of Double Cantilever Beam (DCB) specimens (using cohesive elements for the delamination, in addition to an intralaminar ply failure criterion) were carried out to choose the best interface orientations and laminate configurations. The objective was to ensure a smooth and stable propagation of the interlaminar crack, while preventing asymmetrical delamination growth and plane migration, also known as crack jumping. Then, the best three specimen configurations were manufactured and tested experimentally to validate the simulation methodology. The experimental results confirmed that, with the appropriate stacking sequence for the specimen arms, it is possible to characterize the interlaminar fracture toughness in multidirectional 3D-printed composites under mode I. The experimental results also show that both initiation and propagation values of the mode I fracture toughness depend on the interface angles, although a clear tendency could not be established.