Eccentric training is a mechanical loading classically used in clinical environment to rehabilitate patients with tendinopathies. In this context, eccentric training is supposed to alter tendon mechanical properties but interaction with the other components of the muscle-tendon complex remains unclear. The aim of this study was to determine the specific effects of 14 wk of eccentric training on muscle and tendon mechanical properties assessed in active and passive conditions in vivo. Twenty-four subjects were randomly divided into a trained group (n = 11) and a control group (n = 13). Stiffness of the active and passive parts of the series elastic component of plantar flexors were determined using a fast stretch during submaximal isometric contraction, Achilles tendon stiffness and dissipative properties were assessed during isometric plantar flexion, and passive stiffness of gastrocnemii muscles and Achilles tendon were determined using ultrasonography while ankle joint was passively moved. A significant decrease in the active part of the series elastic component stiffness was found (P < 0.05). In contrast, a significant increase in Achilles tendon stiffness determined under passive conditions was observed (P < 0.05). No significant change in triceps surae muscles and Achilles tendon geometrical parameters was shown (P > 0.05). Specific changes in muscle and tendon involved in plantar flexion are mainly due to changes in intrinsic mechanical properties of muscle and tendon tissues. Specific assessment of both Achilles tendon and plantar flexor muscles allowed a better understanding of the functional behavior of the muscle-tendon complex and its adaptation to eccentric training.