Cell polarity plays key roles in tissue development, regeneration, and pathological processes. However, how the cells establish and maintain polarity is still obscure so far. In this study, by employing microfluidic techniques, we explored the influence of geometrical confinement and chemical stimulation on the cell polarity and their interplay. We found that teardrop shape-induced anterior/posterior polarization of cells displayed homogeneous distribution of epidermal growth factor receptor, and the polarity could be maintained in a uniform epidermal growth factor (EGF) solution, but be broken by a reverse gradient of EGF, implying different mechanism of geometrical and chemical cue-induced cell polarity. Further studies indicated that a teardrop pattern could cause polarized distribution of microtubule-organization center and nucleus-Golgi complex, and this polarity was weakened when the cells were released from the confinement. Our study provides the evidence regarding the difference between geometrical and chemical cue-induced cell polarity and would be useful for understanding relationship between polarity and directional migration of cells.