We have developed a novel magnetic micropillar substrate made of poly(dimethylsiloxane) to apply the mechanical stimulation to individual cell adhesion sites of cells, and to measure associated internal tension of cells. We have successfully embedded enough amount of iron particles into the micropillars and controlled their location on the substrate. By applying magnetic field to the magnetic micropillars, external force can be transferred to the cell adhesion sites formed on the top of these pillars. We investigated change in the internal tension of aortic smooth muscle cells before and after mechanical stimulation using the magnetic pillars. We found that the internal tension of smooth muscle cells fluctuated markedly during releasing of their pretension, which may be due to the tensional homeostasis of smooth muscle cells.