In vitro differentiation of functional human skeletal myotubes in a defined system
In vitro differentiation of functional human skeletal myotubes in a defined system
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DOI：
10.1039/c3bm60166h
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发表时间：
2014-01-01
影响因子：
6.6
通讯作者：
Hickman, James J.
中科院分区：
文献类型：
--
作者：
Guo, Xiufang;Greene, Keshel;Hickman, James J.
体外人类骨骼肌系统是研究人类肌肉发育、疾病和治疗的宝贵工具。然而，已发表的体外人体肌肉系统到目前为止只显示出有限的分化能力。高级分化特征，如横纹和收缩性，只在与运动神经元共培养中观察到。此外，通常认为培养的人肌管不会自发收缩，并且认为任何收缩都源于神经支配。本研究开发了一种无血清培养系统，其中人类骨骼肌管表现出高级分化。免疫细胞化学、电生理和收缩功能分析揭示了这些主要特征：(A)明确的肌肉发育，如交叉条纹的存在所示。(B)发育良好的兴奋-收缩耦合装置，其特征是t小管上的二氢吡啶受体和肌浆网膜上的红嘌呤受体紧密结合。(C)自发和电气控制的收缩力。该报告不仅证明了培养的人骨骼肌管分化水平的提高，而且提供了首次发表的证据，证明这种肌管能够自发收缩。使用这种体外功能的人类骨骼肌系统将推进有关人类骨骼肌发育和生理，以及肌肉相关疾病和治疗的研究。
In vitro human skeletal muscle systems are valuable tools for the study of human muscular development, disease and treatment. However, published in vitro human muscle systems have so far only demonstrated limited differentiation capacities. Advanced differentiation features such as cross-striations and contractility have only been observed in co-cultures with motoneurons. Furthermore, it is commonly regarded that cultured human myotubes do not spontaneously contract, and any contraction has been considered to originate from innervation. This study developed a serum-free culture system in which human skeletal myotubes demonstrated advanced differentiation. Characterization by immunocytochemistry, electrophysiology and analysis of contractile function revealed these major features: (A) Well defined sarcomeric development, as demonstrated by the presence of cross-striations. (B) Finely developed excitation-contraction coupling apparatus characterized by the close apposition of dihydropyridine receptors on T-tubules and ryanodine receptors on sarcoplasmic reticulum membranes. (C) Spontaneous and electrically controlled contractility. This report not only demonstrates an improved level of differentiation of cultured human skeletal myotubes, but also provides the first published evidence that such myotubes are capable of spontaneous contraction. Use of this functional in vitro human skeletal muscle system would advance studies concerning human skeletal muscle development and physiology, as well as muscle-related disease and therapy.