Objective To investigate the synergistic effects of pathologically elevated cyclic stretch and platelet-derived microvesicles (PMVs) on migration of vascular smooth muscle cells (VSMCs) and the potential role of calcium in this process. Methods The FX-5000T cell strain loading system was applied to VSMCs with magnitude of 5% and 15% cyclic stretch to simulate physiological and hypertensive situation respectively in vitro; Wound healing assay was used to analyze the migration of VSMCs; Ca2+-free medium was used to remove extracellular calcium; 2-APB (an antagonist of IP3R) was used to inhibit the release of intercellular stored calcium; GSK219 (an antagonist of TRPV4), and Nifedipine (an inhibitor of L-type voltage-gated calcium channel) were applied to block the activity of respective calcium channel; Thrombin was used to stimulate platelets in vitro which mimics the hypertensive activation of PMVs in vivo. Results Compared with 5% cyclic stretch, 15% cyclic stretch significantly promoted VSMC migration. Removal of extracellular calcium inhibited the migration of VSMCs, but the application of GSK219 and Nifedipine did not affect the migration upregulated by 15% cyclic stretch; while 2-APB which inhibited the release of intracellular stored calcium could also repress the migration of VSMCs during 15% cyclic stretch application. PMVs further promoted VSMC migration under 15% cyclic stretch condition, and both extracellular calcium and intercellular stored calcium were involved in this process. Conclusion The research revealed that both intracellular and extracellular calcium play important roles in VSMC migration induced by 15% cyclic stretch, and PMVs synergistically participate in the above process. The study is aim to provide new mechanobiological insights into the molecular mechanism and clinical targets of vascular remodeling in hypertension.