A microfluidic device with unidirectional perfusion has been developed to observe the effect of human embryonic germ (hEG) cells on SKOV3 cells. The hEG and SKOV3 cells were seeded in the inlet and the outlet reservoirs separately, and co-cultured for 2 days. The medium was perfused unidirectionally from the inlet to the outlet. The growth inhibition of SKOV3 cells was monitored online and the apoptosis signals in SKOV3 culture area decreased along the flow of the medium. In conclusion, microfluidic chip is a potentially useful tool to investigate the effect of stem cells on cancer cells with intuitionistic cell-based screens.