| Expression and Analysis of Flow-regulated Ion Channels in Xenopus Oocytes
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Author:
Date:
2017-04-20
[Abstract] Mechanically-gated ion channels play key roles in mechanotransduction, a process that translates physical forces into biological signals. Epithelial and endothelial cells are exposed to laminar shear stress (LSS), a tangential force exerted by flowing fluids against the wall of vessels and epithelia. The protocol outlined herein has been used to examine the response of ion channels expressed in Xenopus oocytes to LSS (Hoger et al., 2002; Carattino et al., 2004; Shi et al., 2006). The Xenopus oocyte is a reliable system that allows for the expression and chemical modification of ion channels and regulatory proteins (George et al., 1989; Palmer et al., 1990; Sheng et al., 2001; Carattino et al., 2003). ...
[摘要] 机械门控离子通道在机械传导中起关键作用，这是将物理力量转化为生物信号的过程。 上皮细胞和内皮细胞暴露于层流剪切应力（LSS），这是通过流体流向血管壁和上皮细胞壁所施加的切向力。 本文概述的方案已用于检查在非洲爪蟾卵母细胞中表达的离子通道对LSS的反应（Hoger等，2002; Carattino等，2004; Shi等，2006）。 非洲爪蟾卵母细胞是允许离子通道和调节蛋白的表达和化学修饰的可靠系统（George等，1989; Palmer等，1990; Sheng等，2001; Carattino等，2003）。 因此，该技术适用于研究允许流激活通道响应LSS的分子机制。
【背景】排列血管的泌尿道和内皮细胞的上皮细胞经受移动流体引起的机械力。这些力是层流剪切应力（LSS），与管状结构壁相切的摩擦力，以及垂直于流动方向的周向拉伸。令人信服的证据表明，LSS是响应肾和血管管状结构的流动变化而观察到的生理反应的主要决定因素（Satlin等，2001; Liu等，2003; Weinbaum等，2010） 。在这些设置中，离子通道具有将流体剪切应力传递到生物信号中的重要作用（Ranade等，2015）。例如，在肾的远端肾单位中，Na +再吸收和K ...
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| Glioma Associated Stem Cells (GASCs) Isolation and Culture
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Author:
Date:
2015-05-05
[Abstract] Glioma Associated Stem Cells (GASCs) represent a population of non-tumorigenic multipotent stem cells hosted in the microenvironment of human gliomas. In vitro, these cells are able, through the release of exosomes, to increase the biological aggressiveness of glioma-initiating cells. The clinical importance of this finding is supported by the strong prognostic value associated with the GASCs surface immunophenotype thus suggesting that this patient-based approach can provide a groundbreaking method to predict prognosis and to exploit novel strategies that target the tumor stroma (Bourkoula et al., 2014).
[摘要] 胶质瘤相关干细胞（GASC）代表在人类神经胶质瘤的微环境中存在的非致瘤性多能干细胞群体。 在体外，这些细胞能够通过外来体的释放增加胶质瘤起始细胞的生物攻击性。 该发现的临床重要性得到与GASCs表面免疫表型相关的强预后价值的支持，因此表明这种基于患者的方法可以提供开创性的方法来预测预后并开发靶向肿瘤基质的新策略（Bourkoula等人， et al。，2014）。
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| Preparation of Adult Mouse Muscle Stem Cells
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Author:
Date:
2013-12-20
[Abstract] Muscle stem cells are adult stem cells responsible for muscle development, growth and regeneration. Current knowledge suggests those cells are heterogeneous population shared their position between the sarcolemma and basal lamina of muscle fibers. This protocol describes the technique to dissociate and collect the stem cells from skeletal muscle of adult mice, and separate them from other cells found in muscle (e.g. fat, connective tissue). To purify and preserve those myofiber associated muscle stem cells, we use two steps of enzyme digestion followed by cell pre-plating procedures.
[摘要] 肌肉干细胞是负责肌肉发育，生长和再生的成体干细胞。 目前的知识表明这些细胞是异质群体共享他们在肌纤维和肌肉纤维的基底层之间的位置。 该方案描述了从成年小鼠的骨骼肌中分离和收集干细胞并将它们与在肌肉（例如脂肪，结缔组织）中发现的其他细胞分离的技术。 为了纯化和保存这些肌纤维相关的肌肉干细胞，我们使用两个步骤的酶消化，然后细胞预先铺板程序。
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