Myogenesis requires energy production for the execution of a number of regulatory and biosynthesis events. Recent studies in human have shown that mitochondria play a role in the regulation of myogenesis. Indeed, the abundance, morphology, and functional properties of mitochondria contribute to satellite stem cell fate decision during quiescence, proliferation or differentiation. Nevertheless, little attention has been paid to this process in farm animals. Understanding how mitochondria are involved in myogenesis would provide a valuable insight into the underlying mechanisms that regulate the maintenance of cellular homeostasis. The objective of the current study was to evaluate mitochondrial activity including autophagy in satellite cells (SC) during differentiation. Cells were isolated from samples of longissimus and semimembranosus muscle obtained from 4-day-old piglets. Part of the isolated cells was used to separate slow and fast proliferating SC subpopulations by Percoll gradient centrifugation. Both subpopulations were plated in growth medium, allowed to proliferate up to 80% confluence and then placed in an appropriate culture medium to differentiate into myotubes. Mitochondrial function was explored during proliferation and differentiation using Mitotracker dye in flow cytometry. Within the two subpopulations of cells, we observed two different Mitotracker staining intensity suggesting a difference in the mitochondrial membrane potential. According to two distinct peaks of fluorescence within each subpopulation, cells will be separated by cell sorting. Mitochondrial activity will be studied in myogenic cells as well their potential of differentiation. Mitochondria-related proteins expression (TOM20, MTFN1, COXIV) will be also quantified by Western blotting. This study combined with our previous results suggests that autophagy signalling pathways and mitochondria would drive muscle stem cell fate.