We investigated the synergistic effect of D-glucose and epidermal growth factor (EGF) display on the dynamic cellular behaviors of morphology and migration in a culture of human epithelial cells. The time-lapse observation revealed that the cells on the D-glucose/EGF-displayed substrate were endowed with enhanced migration, accompanied with periodic changes in morphology between round and stretched shapes. Immunofluorescence staining of phosphotyrosine PY20 and vinculin was conducted to determine the intracellular localization of phosphorylated tyrosine expression and focal contact formation, respectively. On the substrate displaying D-glucose and EGF, the cells exhibited increases in the levels of the expression of phosphorylated tyrosine and the formation of focal contacts not only at the cellular periphery but also in the cell body. These findings supported the consideration that the displayed D-glucose causes the cells to be in close contact with the surface via grasping glucose transporters on the cytoplasmic membrane.