In this study, we aimed to investigate alterations in secreted proteins as well as expression profile changes in cellular proteome in human skeletal muscle cells that are triggered by a well-established in vitro model of exercise using electrical pulse stimulation (EPS). Here, skeletal muscle cells isolated from muscle biopsies from seven donors were used. The cells from each donor were then exposed to prolonged EPS (i.e. 10 V, 2 ms, 0.1 Hz for 24 h), and non-stimulated cells from the same donors were used as a control.