The goal of this research is to establish a reliable methodology for accelerated fatigue tests of prosthetic heart valves. A polymer valve was the subject, and the influence of various drive parameters on durability was investigated in three different machines. Valve lifetime was notably shortened by increasing the cyclic rate or stroke even though the maximum pressure difference at valve closure was maintained at 120 mm Hg. These results demonstrate that adjustment of the maximum transvalvular pressure is not sufficient to ensure tests are conducted under the same conditions and indicate that measurement of the dynamic load would be more efficacious. Moreover, the locations of tears sustained in the accelerated tests differed from those encountered in an animal experiment although in both cases the locations were entirely consistent with the areas of strain concentration revealed by finite element analysis. These findings should be discussed during a revision of ISO 5840.