Dynamic shifts in metabolic demand during myogenic progression in porcine skeletal muscle stem cells.

Optimizing the metabolic environment during porcine skeletal muscle stem cells (PSCs) proliferation and differentiation is essential for efficient muscle cell cultivation. This study investigated metabolic shifts during myogenic progression using cells at P1, P3, D1, and D2 for characterization and metabolomic analysis. Cell number increased until P3, and myotubes formed by D2 but detached by D3 in 2D culture. Gene and protein expression analyses confirmed myogenic progression, including increased MYOD and MHC expression. Metabolomic profiling identified 94 intracellular metabolites, with 24 and 17 differentially abundant metabolites (DAMs) in proliferation and differentiation, respectively. Pathway analysis showed that energy production via amino acid metabolism was key to progression. Notably, PSCs relied on glycolysis during proliferation and shifted to mitochondrial oxidative phosphorylation during differentiation. These results highlight stage-specific metabolic reprogramming and provide insights for optimizing culture conditions, offering foundational knowledge to improve the efficiency and quality of cultured meat production.