The preparation of carboxymethylated microfibrillated cellulose (MFC) films by dispersion-casting from aqueous dispersions and by surface coating on base papers is described. The oxygen permeability of MFC films were studied at different relative humidity (RH). At low RH (0%), the MFC films showed very low oxygen permeability as compared with films prepared from plasticized starch, whey protein and arabinoxylan and values in the same range as that of conventional synthetic films, e.g., ethylene vinyl alcohol. At higher RH’s, the oxygen permeability increased exponentially, presumably due to the plasticizing and swelling of the carboxymethylated nanofibers by water molecules. The effect of moisture on the barrier and mechanical properties of the films was further studied using water vapor sorption isotherms and by humidity scans in dynamic mechanical analysis. The influences of the degree of nanofibrillation/dispersion on the microstructure and optical properties of the films were evaluated by field-emission scanning electron microscopy (FE-SEM) and light transmittance measurements, respectively. FE-SEM micrographs showed that the MFC films consisted of randomly assembled nanofibers with a thickness of 5-10 nm, although some larger aggregates were also formed. The use of MFC as surface coating on various base papers considerably reduced the air permeability. Environmental scanning electron microscopy (E-SEM) micrographs indicated that the MFC layer reduced sheet porosity, i.e., the dense structure formed by the nanofibers resulted in superior oil barrier properties.