Mangosteen, a fruit mainly produced in Southeast Asia, has been used as food and as an antipyretic and for treating skin diseases. The xanthones contained in mangosteen have many physiological activities including melanin suppression and anticancer activities, but little is known about the physiological effects of the most abundant xanthone, α-mangostin (α-MG) on myoblasts. In this study, we applied α-MG to C2C12 cells that had been induced to differentiate using 2% HS, and analyzed the physiological action of α-MS and the underlying mechanism in the context of myogenic differentiation. α-MG increased the survival rate of C2C12 cells in a concentration-dependent manner. Analysis of the morphological changes in the cells showed that α-MG significantly enhanced the myogenic differentiation of C2C12 myoblasts, whereas the mitochondrial number was only slightly affected. Expression analysis of differentiation-related proteins in C2C12 cells revealed that α-MG promoted the expression of MyoD and Myogenin. Thus, the present study revealed that α-MG improves the survival and myogenic differentiation of C2C12 myoblasts. •α-Mangostin (α-MG) increased C2C12 cell survival.•α-MG significantly enhanced the myogenic differentiation of C2C12 myoblasts.•α-MG promoted the expression of MyoD and Myogenin proteins.