Some different membranes were prepared by Chitosan with the degree of deacetylation (DD) of 63.7%, 73.7%, 83% and 97% respectively. To study the biocompatibility of Chitosan membrane toward corneal stromal cells, the rabbit cells were cultured on the surface of different DD chitosan membranes. The morphological characteristics, the cell-adhesion, the cell proliferation and the activity of LDH in the medium were investigated. The results of experiment shows that the DD of Chitosan has very significant effect on the biocompatibility of Chitosan membrane toward corneal stromal cells. The more DD of Chitosan, the less injury was made to corneal stromal cells by the chitosan membrane, which is favor of the growing and adhesion of corneal stromal cells. The biocompatibility of the membrane made with low DD Chitosan with corneal stromal cells became worse.