An in vitro Mechanical Damage Model of Isolated Myofibers in a Floating Culture Condition.
An in vitro Mechanical Damage Model of Isolated Myofibers in a Floating Culture Condition.
复制标题
漂浮培养条件下分离肌纤维的体外机械损伤模型。
作者：
Y. Tsuchiya;Y. Ono
位于肌纤维表面的肌肉干细胞（卫星细胞）在骨骼肌损伤后从静止状态迅速激活。虽然卫星细胞激活是肌肉再生的第一步，但肌肉损伤对卫星细胞激活的刺激仍有待阐明。我们最近建立了一个体外肌纤维力学损伤模型，分析了从小鼠指长伸肌分离的与肌纤维相关的静止和激活的卫星细胞。在这里，我们描述了一种肌纤维机械损伤的方案，以及在漂浮状态下将完整的健康肌纤维与受损的肌纤维共同培养。这种体外肌纤维损伤模型使我们能够研究不受间质细胞（如血管细胞和成纤维细胞）污染的卫星细胞激活机制，并了解受损的肌纤维衍生因子（DMDFs）如何激活卫星细胞。
Muscle stem cells (satellite cells), located on the surface of myofibers, are rapidly activated from a quiescent state following skeletal muscle injury. Although satellite cell activation is an initial step in muscle regeneration, the stimulation of satellite cell activation by muscle injury remains to be elucidated. We recently established an in vitro mechanical damage model of myofibers, to analyze quiescent and activated satellite cells associated with myofibers isolated from the extensor digitorum longus muscle in mice. Here, we described a protocol for the mechanical damage of myofibers and co-culture of intact healthy myofibers with damaged myofibers in a floating condition. This in vitro myofiber damage model allowed us to investigate the mechanism of satellite cell activation without contamination by interstitial cells, such as blood vessel cells and fibroblasts, as well as understand how damaged myofiber-derived factors (DMDFs) activate satellite cells.