Agricultural by-products, employed as bio-based fillers, provide opportunities to develop new packaging alternatives based on biopolymers. In this study, carrot fibers (CF) and microcrystalline cellulose (MCC) were incorporated into chitosan (CH)-based films at different levels (0-5%, w/w). The obtained films were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, water vapor permeability (WVP), mechanical, thermal, and optical properties. The tensile properties and thermal stability of CH films were improved with the increasing amounts of the fillers. Besides, the higher concentrations of CF reduced the WVP of CH films, whereas 5% MCC addition increased WVP (p < 0.05). The inclusion of both filler type significantly affected the optical properties of CH films. SEM results presented that the fillers were well dispersed throughout the film. Furthermore, the possible interactions between amino groups of CH and functional groups of cellulosic fillers were evaluated by FTIR results. In this study, the potential of CF as filler in producing biopolymer-based packaging materials was investigated.