Enhancing cultured meat production with ginseng leaf-stem extract: a novel supplementation approach to promote porcine muscle stem cell growth

Ginsenosides, including ginsenoside Re (G-Re), exert muscle-protective effects, and ginseng leaves and stems are rich in G-Re. Cultured meat has garnered attention as a sustainable alternative to conventional meat production methods. The proliferation and differentiation of muscle satellite cells (MuSCs) are essential for meat production. Herein, we examined the effects of ginseng leaf-stem extract (G-LSE) on the growth, differentiation, and myotube formation of MuSCs. The G-Re content of G-LSE was quantified using LC-MS/MS. MuSCs were treated with G-LSE (0, 0.25, 0.5, 1, 2, and 4 mg/mL for 48 h). Cell proliferation and cell cycle analyses were performed. Myogenic factors, the AKT/mammalian target of rapamycin (mTOR) signaling pathway, and FoxO3a were detected using real-time PCR, Western blotting, and immunofluorescence analyses. At concentrations of 0.25, 0.5, and 1 mg/mL, G-LSE enhanced cell proliferation over 7 days in a concentration-dependent manner. G-LSE also promoted cell cycle progression. Treatment with G-LSE increased mRNA levels of myogenic differentiation 1 (MyoD), myogenin (MyoG), and myosin heavy chain after 7 days in a concentration-dependent manner. G-LSE also elevated cellular protein levels of MyoD and MyoG. A 7-day treatment with 1 mg/mL G-LSE maintained high MyoG levels and morphological transition to myotubes. Myogenic proliferation and differentiation were mediated via AKT/mTOR activation and inhibition of FoxO3a nuclear translocation. G-LSE promotes porcine MuSC proliferation and differentiation by activating AKT/mTOR signaling and inhibiting FoxO3a nuclear translocation. G-LSE could be a novel plant-derived supplement for promoting MuSC growth and differentiation, particularly in cultured meat production.