Natural extracellular matrices (ECM) provide a more accurate simulation of the cellular growth environment, making them excellent substrate materials for in vitro cell culture. The porcine small intestinal submucosa (SIS) is one of the most widely used natural ECM that display superior bioactivity. However, decellularization operations often result in fiber breakage and failure to recover mechanical strength in the SIS. In the current study, 2,3-dialdehyde cellulose (DAC) was synthesized and cross-linked with SIS gel to form hydrogels. The introduction of DAC into the SIS matrix resulted in a tunable increase in stiffness, which was instrumental in promoting stem cell adhesion and spreading, crucial factors for osteogenic differentiation. The cytotoxicity assessment confirmed the biocompatibility of the SIS-DAC hydrogels indicating their suitability for prolonged cell culture. Moreover, the degradation rate of the hydrogel could be effectively controlled by adjusting the DAC content, addressing the rapid degradation issue associated with SIS gels. This work confirmed the feasibility of using cellulose derivatives to modulate the mechanical properties of matrix gels and influence cell differentiation which offers a valuable experimental foundation for the development of advanced matrix gels tailored for cell culture and regenerative medicine applications.