The small intestinal submucosa (SIS) is an acellular collagen-based matrix, primarily composed of fibrillar collagens (types I, II, and V). They enhance healing due to a minimal immune response. A good degradation rate is the degradation of materials equal to the rate of remodeling in the host. The SIS should apply a good degradation rate and cytocompatibility. In this study, a series of SIS with different degradation rates is obtained by treatment with Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). Morphology, composition, degradable ratio, mechanical properties and cytocompatibility of the SIS are evaluated. We determined a 20μm thickness and 60μm pore size of the native SIS. The degradable ratio of the native SIS was approximately 90% in the presence of 0.25 mg/ml collagenase for 24 hours. The storage modulus of the native SIS was 388MPa. The degradable ratio of the SIS decreased to 6% and the storage modulus increased to 777 MPa after being treated with 30mM EDC for 24 hours. In cytocompatibility assay, cell numbers on the native SIS were similar as on the treated SIS due to the non-toxicity of the EDC treatment process. This SIS exhibited collagenase resistance, stronger mechanical strength and good cytocompatibility after the EDC treatment concluded. The cross-linked SIS could be utilized as a potential cell carrier for tissue engineering application.