The interaction between L929 cells and carboxymethylchitosan (CM-chitosan)-based hydrogels, hydrogels from pure CM-chitosan and its blends, was examined in this study. Cytotoxicity of all materials was also assessed. The cellular morphology and behavior on the surfaces of the hydrogels were observed by scanning electron microscopy (SEM). The effects of various parameters, e.g., type and content of blended polymers, surface structure of hydrogels, and steaming condition used for the preparation of the hydrogels, on the cell-material response were investigated. The results of the cytotoxicity test revealed that all hydrogels were non-cytotoxic. The SEM micrographs demonstrated that the cells proliferated and spread onto a porous CM-chitosan sample. Better cell spreading was found on a flat surface of a CM-chitosan film. Rounded cells were observed when poly(vinyl alcohol) (PVA) was incorporated into CM-chitosan. Fewer cells were found when the content of PVA increased. Spherical clusters of the aggregated cells existed in the blends with ultra high viscosity carboxymethylcellulose (CM-cellulose). In contrast, with the use of low viscosity CM-cellulose, the cells appeared more spreading. The attached cells on the CM-chitosan film steamed at the highest temperature and longest period appeared to spread the most among all tested steaming conditions.