Instrumented gait analysis offers objective clinical outcome assessment. To this purpose, inertial measurement units (IMUs) represent nowadays a very effective solution due to their limited cost, ease of use and improved wearability. The aim of this study was to apply a well-documented IMU-based method to measure gait spatio-temporal parameters in a large number of healthy and gait-impaired subjects, and evaluate its robustness and validity across two clinical centers. Overall, the results of this work represent a robust and reliable foundation for the clinical use of the proposed IMU based method for gait parameters estimation