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首页> 外文期刊>Cell motility and the cytoskeleton >Dissociated flexor digitorum brevis myofiber culture system--a more mature muscle culture system.
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Dissociated flexor digitorum brevis myofiber culture system--a more mature muscle culture system.

机译：离体屈指短肌短肌肌纤维培养系统-一种更成熟的肌肉培养系统。

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Considerable knowledge regarding skeletal muscle physiology and disease has been gleaned from cultured myoblastic cell lines or isolated primary myoblasts. Such muscle cultures can be induced to differentiate into multinucleated myotubes that become striated. However they in general do not fully mature and therefore do not model mature muscle. Contrastingly, fresh and cultured dissociated adult mouse flexor digitorum brevis (FDB) myofibers have been studied for many years. We aimed to investigate the possibility of using the FDB myofiber culture system for drug screening and thus long-term cultures of enzymatically dissociated FDB myofibers were established in 96-well plates. Ca2+ handling experiments were used to investigate the functional state of the myofibers. Imaging of intracellular Ca2+ during electric field stimulation revealed that calcium handling was maintained throughout the culture period of at least 8 days. Western blot and immunostaining analysis showed that the FDB cultures maintained expression of mature proteins throughout the culture period, including alpha-sarcoglycan, dystrophin, fast myosin heavy chain and skeletal muscle alpha-actin. The high levels of the fetal proteins cardiac alpha-actin and utrophin, seen in cultured C2C12 myotubes, were absent in the FDB cultures. The expression of developmentally mature proteins and the absence of fetal proteins, in addition to the maintenance of normal calcium handling, highlights the FDB culture system as a more mature and perhaps more relevant culture system for the study of adult skeletal muscle function. Moreover, it may be a useful system for screening therapeutic agents for the treatment of skeletal muscle disorders.
机译：已从培养的成肌细胞系或分离的原代成肌细胞中收集了有关骨骼肌生理学和疾病的大量知识。可以诱导这种肌肉培养物分化成横纹的多核肌管。但是，它们通常不能完全成熟，因此不能模拟成熟的肌肉。相反，已经研究了新鲜的和培养的离体成年小鼠屈指短肌（FDB）肌纤维。我们旨在研究使用FDB肌纤维培养系统进行药物筛选的可能性，因此在96孔板中建立了酶解FDB肌纤维的长期培养。 Ca2 +处理实验用于研究肌纤维的功能状态。电场刺激过程中细胞内Ca2 +的成像显示，钙的处理在至少8天的整个培养过程中均得以保持。 Western印迹和免疫染色分析表明，FDB培养物在整个培养期间均保持成熟蛋白的表达，包括α-糖聚糖，肌营养不良蛋白，快速肌球蛋白重链和骨骼肌α-肌动蛋白。在FDB培养物中，在培养的C2C12肌管中未见高水平的胎儿蛋白心脏α-肌动蛋白和卵磷脂。除了维持正常的钙处理外，发育成熟蛋白的表达和胎儿蛋白的缺失，突出显示了FDB培养系统是用于研究成人骨骼肌功能的更成熟且也许更相关的培养系统。而且，它可能是筛选用于治疗骨骼肌疾病的治疗剂的有用系统。

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