In this study, we present a role of arrayed micropillar structures in cell rolling dynamics. Cell rolling on a ligand coated surface as a means of cell separation was demonstrated using a micropillar-integrated microfluidic channel. This approach allows the separation of cells according to characteristic surface properties, regardless of cell size. In these experiments, different moving trajectories of the cells between a ligand-coated micropost structure and a 1% BSA coated micropost structure were observed using sequential images. Based on the analysis of the angle of travel of cells in the trajectory, the average angles of travel on the ligand-coated microposts were 1.5° and -3.1° on a 1% BSA-coated micropost structure. The overall force equivalent applied to a cell can be analyzed to predict the cell rolling dynamics when a cell is detached. These results show that it will be possible to design chip geometry for delicate operations and to separate target cells. Furthermore, we believe that these control techniques based on a ligand coated micropillar surface can be used for enhancing cell rolling-based separation in a faster and more continuous manner.