Abstract
Cultured meat, a sustainable alternative to conventional meat, addresses ethical and environmental challenges in livestock production. Its production relies on bovine muscle stem cells from adult muscle or fetal tissue, whose proliferation and differentiation vary with age and developmental stage. However, the molecular mechanisms underlying these variations remain unclear. RNA sequencing was performed to characterize the transcriptomic landscape of bovine muscle stem cells across developmental stages, including myogenic maturation. Differentially expressed genes and key signaling pathways regulating myogenesis were identified, and the functional impact of modulating the AKT-autophagy pathway on differentiation was assessed. Transcriptomic analysis revealed distinct age-dependent gene expression patterns. It was possible to classify cells into three categories: young undifferentiated, young differentiated, and old differentiated. Young undifferentiated-like cells exhibited upregulation of genes associated with active states during the transitions from quiescence to activation and, ultimately, to commitment, indicating that they had robust differentiation potential. In contrast, aged myogenic samples displayed gene expression profiles that acted as barriers to efficient myogenic differentiation. Notably, modulation of the AKT-autophagy pathway both facilitated the production of very mature myogenic cells and prevented spontaneous differentiation, thereby preserving differentiation capacity in vitro. These findings provide insights into age-dependent muscle stem cell differentiation and suggest strategies to optimize cultured meat production. The appropriate modulation of key signaling pathways may help us to overcome major challenges in achieving scalable and efficient cultured meat manufacturing.