| Isolation, Culture, and Staining of Single Myofibers
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Author:
Date:
2016-10-05
[Abstract] Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single ...
[摘要] 成年骨骼肌再生由被称为卫星细胞的成人干细胞的专门群体协调，其被定位在基底层和肌纤维的质膜之间。在肌肉再生期间发生的卫星细胞激活，增殖和随后分化的过程可以通过从骨骼肌中分离单个肌纤维并在悬浮条件下培养它们而在体外重现。在这里，我们描述了一种改进的协议，通过从小鼠的伸肌腱长肌（EDL）肌肉中分离单个肌纤维来评价离体细胞的活化，并培养和染色肌纤维相关的卫星细胞的标记物自我更新，增殖和分化。
[背景] 虽然骨骼肌是一种完全分化的有丝分裂后组织，但是它保持了内在的能力，以对遗传和获得性肌肉纤维损伤的形式Grand和Rudnicki，2007）。成年人的肌肉再生由称为卫星细胞的干细胞群体介导，所述细胞位于有丝分裂静止状态的肌纤维的基底层和肌纤维膜之间（Le Grand和Rudnicki，2007）。响应于肌肉创伤，卫星细胞变得活化和增殖以产生与预先存在的纤维融合的成肌细胞，并彼此相互修复或产生新的肌纤维。一小部分卫星细胞不分化，而是重新进入静止以维持干细胞库。所有哺乳动物物种的卫星细胞表达配对盒（Pax）转录因子Pax7，其也用作确定与其他生肌因子如MyoD相关的卫星细胞命运的关键标记（Le Grand和Rudnicki，2007; et al。，2006; Kuang and Rudnicki，2008）。
可以通过肌纤维外植体的悬浮培养部分地重现关于卫星细胞活化，增殖和分化的肌肉再生的体内过程（Rosenblatt等人，1995; ...
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| Isolation and Primary Cell Culture of Mouse Dorsal Root Ganglion Neurons
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Author:
Date:
2016-04-05
[Abstract] We here provide a detailed protocol for the isolation and culture of primary mouse sensory neurons. The cell bodies of sensory afferent pseudounipolar neurons are located in dorsal root ganglia (DRGs) along the vertebral column. Dissected mouse DRGs can be dissociated into single cells by enzymatic digestion to obtain primary cultures of mouse sensory neurons as performed in the studies reported by Khaminets et al. (2015).
[摘要] 我们在这里提供了详细的协议，用于隔离和培养的主要小鼠感觉神经元。 感觉传入假性极化神经元的细胞体位于沿着脊柱的背根神经节（DRG）中。 解离的小鼠DRG可以通过酶消化解离成单个细胞，以获得小鼠感觉神经元的原代培养物，如Khaminets等人（2015）报道的研究中所进行的。
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