Relative Stiffness Measurements of Cell-embedded Hydrogels by Shear Rheology in vitro
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Author:
Date:
2017-01-05
[Abstract] Hydrogel systems composed of purified extracellular matrix (ECM) components (such as collagen, fibrin, Matrigel, and methylcellulose) are a mainstay of cell and molecular biology research. They are used extensively in many applications including tissue regeneration platforms, studying organ development, and pathological disease models such as cancer. Both the biochemical and biomechanical properties influence cellular and tissue compatibility, and these properties are altered in pathological disease progression (Cox and Erler, 2011; Bonnans et al., 2014). The use of cell-embedded hydrogels in disease models such as cancer, allow the interrogation of cell-induced changes in the biomechanics of the microenvironment (Madsen et al., 2015). Here we report a simple method to ...
[摘要] 由纯化的细胞外基质(ECM)组分(如胶原,纤维蛋白,Matrigel和甲基纤维素)组成的水凝胶系统是细胞和分子生物学研究的支柱。它们广泛用于许多应用,包括组织再生平台,研究器官发育和病理疾病模型如癌症。生物化学和生物力学性质都影响细胞和组织相容性,并且这些性质在病理疾病进展中发生改变(Cox和Erler,2011; Bonnans等人,2014)。在诸如癌症的疾病模型中使用细胞嵌入的水凝胶允许询问细胞诱导的微环境生物力学变化(Madsen等人,2015)。在这里,我们报告一种使用受控应变旋转流变仪测量这些细胞诱导的体外变化的简单方法。
背景 纤维化和实体瘤伴随着其天然组织的病理重塑(Cox和Erler,2011; Bonnans等人,2014)。在两种病理状况下,局部组织环境经历物理化学和生物学变化,导致组织刚度(弹性模量)增加(Humphrey等人,2014)。增强的组织/基质调节导致细胞行为改变,细胞形态,分化状态,增殖,迁移和干性的机械信号。在癌症的临床前动物模型中,这些变化可以驱动恶性进展和转移性扩散(Bonnans等人,2014)。不足为奇的是,靶基质硬化近年来受到了极大的关注,几项临床试验已经开始(Kai等人,2016)。  基质组分的弹性和机械性能可以使用原子力显微镜(AFM)进行检查,原子力显微镜(AFM)是一种提供纳米分辨率并以picoNewton分辨率同时测量施加力的技术(Kasas和Dietler,2008)。然而,AFM不适用于理解较大的3D矩阵的弹性特性。使用剪切流变学可以更精确地检查体积3D矩阵的机械性能(Picout和Ross-Murphy,2003)。流变学是研究当施加力时材料如何变形。因此,将剪切应力施加到3D矩阵可以确定体积3D矩阵的弹性模量(刚度)。在该方案中,我们描述了一种通过剪切流变学测量细胞诱导的与癌相关成纤维细胞嵌入的水凝胶的基质刚度变化的方法。 ...
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Clonal Culture of Mouse Liver Progenitor Cells
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Author:
Date:
2015-10-20
[Abstract] Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. This protocol describes the process of cell isolation and colony assay to examine proliferative and differentiation potential of LPCs.
[摘要] 肝干/祖细胞(LPC)被定义为分化成肝细胞和胆管细胞的双能细胞。 为了分析其分化潜力,克隆培养已经用于通过细胞分选仪分离的LPC。 此外,我们可以使用文化来评估目标基因的功能分化潜力的LPCs。 该协议描述细胞分离和集落分析的过程,以检查增殖和分化潜力的LPCs。
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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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