Nanocrystalline cellulose (NCC) was prepared from microcrystalline cellulose (MCC) with particle size of 24.7 μm using sulfuric acid hydrolysis technique. The obtained NCC revealed size of 0–100 nm, which the major part of them was about 30 nm. Then different contents (2.5, 5 and 10 wt%) of these NCC incorporated in agar film solution and the morphology, structure, and properties of the nanocomposite films were characterized by scanning electron microscope (SEM), X-ray diffraction (XRD), Fourier transforms infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), mechanical, physical and optical testing. Results showed that the water vapor permeability (WVP) and water solubility (WS) of the agar-based nanocomposite films significantly (P < 0.05) decreased about 13% and 21%, respectively, upon increasing the NCC content to 10%. Tensile strength (TS) and Young's modulus (YM) values of nanocomposite films significantly increased (P ≤ 0.05) with addition of NCC, whereas the elongation percent (E%) decreased not significantly (P > 0.05). In addition, swelling percentage, transparency and light transmission of the films were decreased by incorporating NCC into polymer matrix.