In this study, the effects of repetition of assembly and disassembly of polymer electrolyte membrane fuel cells on electrochemical performance were systematically investigated. Additionally, the effects of additional activation on polymer electrolyte membrane fuel cells were evaluated. All fuel cells were measured every three days. For the disassembled polymer electrolyte membrane fuel cells, membrane electrode assemblies were stored in a vacuum desiccator. For the maintained assembly, fuel cells were stored at room temperature. The performance and electrochemical characteristics of the fuel cell were analyzed by electrochemical impedance spectroscopy. As a result, the addition of activation to maintained assembly fuel cells showed the best performance among fuel cells with other assembly and activation conditions. Repetition of assembly and disassembly, as well as insufficient activation, caused degradation of the performance of fuel cells.