Isolation, culture, and use of primary murine myoblasts in small-molecule screens

Small-molecule screens (SMS) are often performed using transformed cell lines that have limited physiological relevance to the biological system being investigated, resulting in poor translational outcomes. To circumvent this limitation, we present a protocol to perform SMS in primary murine myoblasts. We describe steps for isolating primary skeletal muscle myoblasts with greater than 95% purity, then describe techniques to establish a robust dynamic range, and conclude with steps to initiate a successful SMS. For complete details on the use and execution of this protocol, please refer to Richler and Yaffe (1970),¹ Rando and Blau (1994),² and Earle et al. (2020).³.