Vascular smooth muscle cells (VSMCs) are constantly exposed to cyclic stretch in the body, which makes it beneficial to study the effects of cyclic stretch on VSMCs. In this study, we developed a poly(dimethyl siloxane) (PDMS) compact six-well device that can be used to study the combined effect of cyclic strain and various growth factors on cultured VSMCs. Cell adhesion, alignment, and proliferation under 10% or 20% cyclic strain at 1 Hz were studied using this surface-enhanced PDMS device. The combined effects of cyclic strain with either transforming growth factor-, vascular endothelial growth factor, fibroblast growth factor, or epidermal growth factor on VSMC proliferation was also examined. Results showed that VSMCs adhered well on the surface-enhanced multiwell device and they aligned perpendicularly to the direction of the cyclic strain. Cell proliferation was inhibited by 10% cyclic strain at 1 Hz compared with static control. The mitogenic effects of the growth factor were less potent under either 10% or 20% cyclic strain. With simple modification to accommodate more wells, this device could potentially be a useful tool for more economical, high throughput screening application.