A new hydroxylpropyl chitosan (HPCTO) capable of forming both thermotropic and lyotropic liquid crystalline phases was synthesized by reaction of alkali chitosan with propylene oxide and its solid films cross-linked with glyoxal were prepared by casting the liquid crystalline solution in methanol. The thermal and swelling properties of the cross-linked films were investigated. The films displayed fingerprint patterns characteristic of cholesteric liquid-crystalline phase, and their pitches increased with increasing temperature and cross-linker concentration. The cross-linked samples exhibited an anisotropic swelling in both water and methanol, suggesting that the two-dimensional cross-linking preferentially performs between HPCTO molecules. The degree of anisotropy highly depended on the solvent, but hardly on the cross-linker concentration investigated.