Simulations of cardiac electrophysiology and mechanics have been reported to be sensitive to the microstructural anisotropy of the myocardium. Consequently, a personalized representation of cardiac microstructure is a crucial component of accurate, personalized cardiac biomechanical models. In-vivo cardiac Diffusion Tensor Imaging (cDTI) is a non-invasive magnetic resonance imaging technique capable of probing the heart’s microstructure. Being a rather novel technique, issues such as low resolution, signal-to noise ratio, and spatial coverage are currently limiting factors. We outline four interpolation techniques with varying degrees of data fidelity, different amounts of smoothing strength, and varying representation error to bridge the gap between the sparse in-vivo data and the model, requiring a 3D representation of microstructure across the myocardium. We provide a workflow to incorporate in-vivo myofiber orientation into a left ventricular model and demonstrate that personalized modelling based on fiber orientations from in-vivo cDTI data is feasible. The interpolation error is correlated with a trend in personalized parameters and simulated physiological parameters, strains, and ventricular twist. This trend in simulation results is consistent across material parameter settings and therefore corresponds to a bias introduced by the interpolation method. This study suggests that using a tensor interpolation approach to personalize microstructure with in-vivo cDTI data, reduces the fiber uncertainty and thereby the bias in the simulation results.