Porous scaffold is widely used in the field of tissue engineering. However, the anisotropic structure of actual extracellular matrix (ECM) of human tissue pose a challenge to the scaffold structure that pore size should be changed in gradient. Here we report a stage cooling method to fabricate alginate scaffold with gradient pores. Eight cooling models were set according to different temperature steps, different initial temperature, and different time duration. The thermal characterization of solution during cooling process were recorded and scaffold morphology were observed. The results revealed that the temperature steps mainly affected pore shape, while the initial temperature and time duration mainly affected pore size. By altering the initial temperature and time duration, scaffold exhibited cellular and gradually enlarged pores on the vertical axial direction (10-65 μm at base, 50-141 μm at top). With this stage cooling method, pore shape and pore size could be easily tailored and scaffold with gradient structure could be fabricated.