Cells are inherently exposed to a number of different biophysical stimuli such as electric fields, shear stress, and tensile or compressive stress from the extracellular environment in vivo. Each of these biophysical cues can work simultaneously or independently to regulate cellular functions and tissue integrity in both physiological and pathological conditions. Thus, it is vital to understand the interaction of multiple stimuli on cells by decoupling and coupling the stimuli in simple combinations and by investigating cellular behaviors in response to these cues. Here, we report a novel microfluidic platform to apply the combinatorial stimulation of an electric field and fluid shear stress by controlling two directional cues independently. An integrated microfluidic platform was developed using soft lithography to monitor the cellular migration in real-time in response to an electric field and fluid shear stress in single, simultaneous, and sequential modes. When each of these stimulations is applied separately, normal human dermal fibroblasts migrate toward the anode and in the direction of fluid flow in a dose-dependent manner. Simultaneous stimulation with an electric field and shear stress, which mimics a wound in vivo, enhances the directional migration of fibroblasts by increasing both directedness and trajectory speed, suggesting the plausible scenario of cooperation between two physical cues to promote wound healing. When an electric field and shear stress are applied sequentially, migration behavior is affected by the applied stimulation as well as pre-existing stimulating conditions. This microfluidic platform can be utilized to understand other microenvironments such as embryogenesis, angiogenesis and tumor metastasis.