Dr. John and Anne Chong Lab for Functional Genomics, Charles Perkins Centre, School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia.
Larance Laboratory, Charles Perkins Centre, School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW 2006, Australia.
Cells. 2022 Mar 11;11(6):963. doi: 10.3390/cells11060963.
Skeletal muscle weakness is linked to many adverse health outcomes. Current research to identify new drugs has often been inconclusive due to lack of adequate cellular models. We previously developed a scalable monolayer system to differentiate human embryonic stem cells (hESCs) into mature skeletal muscle cells (SkMCs) within 26 days without cell sorting or genetic manipulation. Here, building on our previous work, we show that differentiation and fusion of myotubes can be further enhanced using the anabolic factors testosterone (T) and follistatin (F) in combination with a cocktail of myokines (C). Importantly, combined TFC treatment significantly enhanced both the hESC-SkMC fusion index and the expression levels of various skeletal muscle markers, including the motor protein myosin heavy chain (MyHC). Transcriptomic and proteomic analysis revealed oxidative phosphorylation as the most up-regulated pathway, and a significantly higher level of ATP and increased mitochondrial mass were also observed in TFC-treated hESC-SkMCs, suggesting enhanced energy metabolism is coupled with improved muscle differentiation. This cellular model will be a powerful tool for studying in vitro myogenesis and for drug discovery pertaining to further enhancing muscle development or treating muscle diseases.
骨骼肌无力与许多不良健康后果有关。由于缺乏足够的细胞模型，目前识别新药的研究往往没有定论。我们之前开发了一种可扩展的单层系统，可以在 26 天内无需细胞分选或遗传操作将人胚胎干细胞（hESC）分化为成熟的骨骼肌细胞（SkMC）。在这里，基于我们之前的工作，我们表明使用合成代谢因子睾酮（T）和卵泡抑素（F）以及肌因子鸡尾酒（C）可以进一步增强肌管的分化和融合。重要的是，联合 TFC 治疗显著提高了 hESC-SkMC 的融合指数和各种骨骼肌标志物的表达水平，包括运动蛋白肌球蛋白重链（MyHC）。转录组学和蛋白质组学分析显示氧化磷酸化是上调最明显的途径，在 TFC 处理的 hESC-SkMC 中也观察到 ATP 水平显著升高和线粒体质量增加，这表明能量代谢的增强与肌肉分化的改善相关。这种细胞模型将成为研究体外肌发生和药物发现的有力工具，有助于进一步增强肌肉发育或治疗肌肉疾病。
