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

Yue Qu; Kaydine Edwards; Joeva Barrow

doi:10.1016/j.xpro.2023.102149

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

STAR Protoc. 2023 Mar 13;4(2):102149. doi: 10.1016/j.xpro.2023.102149. Online ahead of print.

Authors

Yue Qu¹, Kaydine Edwards¹, Joeva Barrow²

Affiliations

¹ Division of Nutritional Sciences, Cornell University, Ithaca, NY 14850, USA.
² Division of Nutritional Sciences, Cornell University, Ithaca, NY 14850, USA. Electronic address: jb2254@cornell.edu.

Abstract

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.

Keywords: Cell Biology; Cell Culture; Cell Isolation; Cell-based Assays; High-throughput Screening; Metabolism.