Isolated individual myofibers are valuable experimental models that can be used in various conditions to understand skeletal muscle physiology and pathophysiology at the tissue and cellular level. This report details a time- and cost-effective method for isolation of single myofibers from the flexor digitorum brevis (FDB) muscle in both young and aged mice. The FDB muscle was chosen for its documented history in single myofiber experiments. By modifying published methods for FDB myofiber isolation, we have optimized the protocol by first separating FDB muscle into individual bundles before the digestion, followed by optimizing the subsequent digestion medium conditions to ensure reproducibility. Morphological and functional assessments demonstrate a high yield of isolated FDB myofibers with sarcolemma integrity achieved in a shorter time frame than previous published procedures. This method could be also adapted to other types of skeletal muscle. Additionally, this highly reproducible method can greatly reduce the number of animals needed to yield adequate numbers of myofibers for experiments. Thus, this advanced method for myofiber isolation has the potential to accelerate research in skeletal muscle physiology and screening potential therapeutics "ex vivo" for muscle diseases and regeneration.

中文翻译：

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从指屈肌短肌中快速分离单根肌纤维的先进方法。

分离的个体肌纤维是有价值的实验模型，可用于各种条件下以了解组织和细胞水平的骨骼肌生理学和病理生理学。该报告详细介绍了一种从年轻和老年小鼠的指短屈肌 (FDB) 肌肉中分离单根肌纤维的时间和成本有效的方法。选择 FDB 肌肉是因为它在单肌纤维实验中有记录的历史。通过修改已发表的 FDB 肌纤维分离方法，我们首先在消化前将 FDB 肌肉分离成单独的束，然后优化后续消化介质条件以确保重现性，从而优化了协议。形态学和功能评估表明，与之前公布的程序相比，在更短的时间内实现了高产率的分离 FDB 肌纤维和肌膜完整性。该方法也适用于其他类型的骨骼肌。此外，这种高度可重复的方法可以大大减少产生足够数量的实验用肌纤维所需的动物数量。因此，这种先进的肌纤维分离方法有可能加速骨骼肌生理学的研究，并筛选肌肉疾病和再生的“离体”潜在疗法。