Carboxymethyl cellulose (CMC) and glycerol were added by immersing neutralized collagen fiber paste into CMC/glycerol aqueous solutions with different mass ratios, forming composite films. CMC decreased the glycerol adsorption by 49.5%. The presence of CMC formed rougher film surface and more compact internal structure. Glycerol increased the distances between collagen molecules and decreased the relative triple helix contents. The tensile strength and Young’s modulus of film in dry condition maximum respectively increased by 125% and 277% with the immersion in CMC (1.5%)/glycerol (4%) solution, compared to the sample immersed into pure glycerol solution, but mechanical properties decreased at high glycerol content. Besides, lower wet mechanical strength was observed as the addition of CMC, corresponding to the higher water swelling ratio of film. CMC also improved the shrinkage stability under boiling and the thermostability of film at a fixed glycerol level, but higher glycerol content caused the decrease of denaturation temperature of film.