In this study, independent control of mechanical and diffusional properties of hydrogel could be achieved by employing polymeric crosslinker that allows for the adjustment of the number of crosslinking sites per molecule (degree of substitution). By using the polymeric crosslinker, with varying degree of substitution, the moduli of the resulting hydrogels could be controlled in a wide range without changing their concentration. Furthermore, their diffusional properties, as characterized by their swelling ratios, pore diameters and drug release rates, were not significantly affected by the changes in degree of substitution. The viability of cells encapsulated in this hydrogel system successfully demonstrated the varying effects of mechanical properties on different cell types, whereas the cells in a conventional hydrogel system were more significantly influenced by changes in diffusion.