Simplifying the protocol for low-pollution-risk, efficient mouse myoblast isolation and differentiation

Summary Myoblasts are the primary effector cells that play crucial roles in myogenesis and muscle regeneration following injury. However, isolating purified primary myoblasts from murine skeletal muscle poses challenges for junior researchers. Here, we present a simplified, low-risk, and optimized protocol for the extraction and enrichment of these myogenic progenitor cells. Additionally, we demonstrate that, compared to F10 (Ham’s F-10)-based medium, DMEM (Dulbecco’s Modified Eagle’s Medium)-based differentiation medium provides a more conducive environment for myoblasts differentiation. This enhancement improves the efficiency of myofiber formation and the expression of myogenic markers. Graphical Abstract A convenient, low-pollution-risk, and expeditious methodology for isolating primary myoblasts has been developed. Comparative analysis of myoblast differentiation under DMEM and F10-based medium conditions revealed that myotubes formed in DMEM displayed a more robust and hypertrophic phenotype.