For the first time, we have developed a microfluidic device for on-chip monitoring of suspension cell-cell communication from stimulated to recipient HL-60 cells. A deformable PDMS membrane was developed as a compressive component to perform cell entrapment and exert different modes of mechanical stimulation. The number of cells trapped by this component could be modulated by flushing excessive cells towards the device outlet. The trapped cells could be triggered to release mediators by mechanical stimulation. Sandbag microstructures were used to immobilize recipient cells at well-defined positions. These recipient cells were evoked by mediators released from mechanically stimulated cells trapped in the compressive component. Normally closed microvalves were integrated to provide continuous-flow and static environment. We studied cell-cell communication between stimulated (in compressive component) and recipient (in sandbag structures) cells. Calcium oscillations were observed in some recipient cells only when a low number of cells were stimulated. Different mechanical stimulation and flow environment were also employed to study their impact on the behavior of cell-cell communication. We observed that both the duration and intensity of intracellular calcium responses increased in persistent stimulation and decreased in flowing environment. This microdevice may open up new avenues for real-time monitoring of suspension cell-cell communication, which propagates via gap-junction independent mechanism, with multiple variables under control.