The cathode degradation of a direct methanol fuel cell (DMFC) was investigated after a 240 h discontinuous galvostatic operation at 80 °C. The catalyst coated membrane (CCM) and the cathode diffusion layer were not combined so as to isolate electrochemical and mass transport processes. It was indicated by the EDS and SEM tests that the loss of the cathode electrochemical surface area (ESA) was associated with the decays of the Pt/C catalyst and the interfacial contact. Furthermore, Ru crossover and higher methanol crossover resulting from the anode failure aggravated the degradation of the cathode. On the other hand, the change of the pore structure led to a higher wettability of the cathode microporous layer. Therefore, the oxygen transport was suppressed due to the decrease of hydrophobic passages.