Metabolomics-guided isolation of myogenic progenitor cells for improved transplantation

Skeletal muscle maintains considerable capacity for regeneration following injury, but successful regeneration is limited in instances of volumetric muscle loss, advanced aging, or muscular dystrophies. Considerable research has been done on muscle stem cell (MuSC) transplantation; however, proliferative exhaustion and donor cell dose requirements have slowed progress. Due to the paramount role of cellular metabolism in regenerative function of stem cells, the clinical potential for MuSC therapy may be improved by minimizing the isolation-induced metabolic perturbations experienced by MuSCs. This study uses a model of simulated cell sorting combined with untargeted, small-molecule metabolomic profiling to outline sorting-induced metabolic perturbations in C2C12 myoblasts. We expand upon this by performing a time course of metabolomic profiling on myoblasts recovering from either fluorescence-activated cell sorting or magnetic-bead-activated cell sorting-based isolation procedures to determine the method and recovery timing for optimal redox and energetic status. Using this metabolism-informed method, we then performed primary MuSC transplantation studies in mice to demonstrate the generalizability from the in vitro system to in vivo MuSC transplantation during regeneration from barium chloride-induced injury. Our findings demonstrate metabolically favorable strategies to isolate MuSC for analysis of the quiescent-to-activated metabolic transition or enhance transplantation efficacy.NEW & NOTEWORTHY Due to the paramount role of cellular metabolism in regenerative function of stem cells, the clinical potential for muscle stem cell (MuSC) therapy may be improved by minimizing isolation-induced metabolic perturbations. This study uses a model of simulated cell sorting combined with untargeted, small-molecule metabolomic profiling to outline sorting-induced metabolic perturbations in myoblasts. Our findings demonstrate metabolically favorable strategies to isolate MuSC for analysis of the quiescent-to-activated metabolic transition or enhance transplantation efficacy.