Cell detachment is a routine process to recover seeded cells from culture systems. However, traditional enzymatic methods might easily induce senescence or apoptosis of cells during the routine passage process due to an improper handing process. The pH-responsive characteristic of chitosan has been demonstrated to control cell attachment/detachment by precise adjustment of medium pH within 1 h without additional enzymatic treatment and extensive washing steps. But the time 1 h is not satisfying to all applications to recover adherent cells from chitosan surface. The purpose of this study is to regulate pH-responsive characteristics of chitosan by blending with non-pH-responsive biomaterials, polycaprolactone (PCL) and polyvinyl alcohol (PVA) to provide a gentle rapid method for cell detachment. In this study, PCL could delay the pH-responsive characteristics of chitosan to reduce cell detachment ratio (< 65%) after raising medium pH to 7.65 for 2 h. In contrast, PVA could provide a synergistic effect on rapid detachment of cells (>95%) from the chitosan/PVA blended surface within 30 min after raising medium pH to 7.65. We further discuss the related mechanism among cell detachment, fibronectin, and chitosan-based blends. These results indicate non-pH-responsive PCL and PVA could be added into pH-responsive chitosan to develop a rapid non-enzymatic strategy to control cell detachment rate for cell passage and various tissue engineering applications.