In this study, polycaprolactone (PCL) cylindrical scaffolds with gradually increasing pore size along the longitudinal direction were fabricated by the centrifugation and thermal fibril-bonding process. The fabricated PCL scaffold showed a gradual increasing pore size (from approximately 90 to approximately 400 microm) and porosity (from approximately 80 to approximately 97%) along the cylindrical axis. The pore size gradient PCL scaffold was used to investigate the effect of pore size on the chondrogenic differentiation of adipose stem cells (ASCs). From the in vitro culture of ASCs, it was observed that the scaffold section having a pore size range of 370-400 microm provided a more favorable environment for chondrogenic differentiation than other pore size groups. The pore size gradient scaffolds can be a good tool for the systematic study of determining optimum pore size ranges for a variety of stem cell differentiation to a specific cell type.