There are few studies regarding the effects of mechanical stimulation on cell migration although biochemical factors have been widely studied. We have investigated the effects of intermittent hydrostatic pressure (IHP) on mesenchymal stem cell migration with or without neighboring endothelial cells (EC). IHP promoted MSCs migration and the neighboring ECs helped with this. However, when IHP was applied to MSCs cultured with ECs, the opposite effect was observed. The concentration of stromal-derived factor-1 culture in medium was measured to explain the obtained results. SDF-1 concentration increased as IHP increased when MSCs were cultured alone. However, it decreased as IHP increased when MSCs and ECs were co-cultured. These results indicate that the mechanical environment should be considered when studying the migration of a cell type along with its biochemical environment.