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Y-27632 dihydrochloride

Y-27632 dihydrochloride

Company: Sigma-Aldrich
Catalog#: Y0503
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Ex vivo Culture of Adult Mouse Antral Glands
Author:
Date:
2017-01-05
[Abstract]  The tri-dimensional culture, initially described by Sato et al. (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker et al. (2010), who efficiently generated organoids that recapitulate the mature pyloric epithelium in vitro. This ex vivo approach is suitable and promising to study gastric function in homeostasis as well as in disease. We have adapted Barker’s protocol to compare homeostatic and regenerating tissues and here, we meticulously describe, step by step, the isolation and culture of antral glands as well as the isolation of single cells from antral glands that ... [摘要]  为了分离和表征成年小肠的上皮干细胞,最初由Sato等人(2009)描述的三维培养物已经随后适应于许多不同的器官。其中一个例子是Barker等人(2010)分离和培养窦性干细胞,他们有效地产生了在体外重现成熟幽门上皮的组织细胞。这种“离体”方法是适合的,并且有希望研究体内平衡和疾病中的胃功能。我们已经调整了Barker的方案来比较稳态和再生组织,这里,我们一步一步地仔细地描述了窦腺的分离和培养,以及从细胞分选后可能用于培养的窦腺中分离单细胞一个例子(Fernandez Vallone等人,2016)。

背景来自腺体的小鼠成体干细胞可以在3D matrigel中离体生长,作为“迷你腺体”无限期(Barker等人,2010) 。与EGF,Noggin和R-spondin 1存在下生长的小鼠成年小肠的干细胞相比,胃干细胞需要进一步补充Fgf10,胃泌素,Wnt3a和更高浓度的R-螺旋菌素1(称为作为ENRFGW)获得生产性文化。直到最近,在干细胞消融后,在离体培养系统中成体再生窦腺是否生长,如果是这样,仍然是未知的。使用本方案,证明了内源性和再生的腺体在接种时不会类似地生长并且表现出不同的生长培养要求。

Ex vivo Culture of Fetal Mouse Gastric Epithelial Progenitors
Author:
Date:
2017-01-05
[Abstract]  Isolation and tridimensional culture of murine fetal progenitors from the digestive tract represents a new approach to study the nature and the biological characteristics of these epithelial cells that are present before the onset of the cytodifferentiation process during development. In 2013, Mustata et al. described the isolation of intestinal fetal progenitors growing as spheroids in the ex vivo culture system initially implemented by Sato et al. (2009) to grow adult intestinal stem cells. Noteworthy, fetal-derived spheroids have high self-renewal capacity making easy their indefinite maintenance in culture. Here, we report an adapted protocol for isolation and ex vivo culture and maintenance of fetal epithelial progenitors from distal pre-glandular ... [摘要]  来自消化道的鼠胎儿祖细胞的分离和三维培养代表了研究在发育过程中细胞分化过程开始前存在的这些上皮细胞的性质和生物学特征的新方法。在2013年,Mustata等人描述了在最初由佐藤等人实施的离体培养系统中分离作为球体生长的肠胎细胞祖细胞。 >(2009)增长成年肠干细胞。值得注意的是,胎儿衍生的球体具有较高的自我更新能力,使其在文化中的无限期维护变得容易。在这里,我们报告了用于分离和远离前胃腺胃生长为胃球体的胎儿上皮祖细胞的分离和离体培养和维持的修改方案(Fernandez Vallone等人, 2016)。

背景 来自腺体的小鼠成体干细胞可以在3D matrigel中离体生长,作为“迷你腺体”无限期(Barker等人,2010) 。与在EGF,Noggin和R-spondin 1存在下生长的小肠的干细胞相比,成年胃干细胞需要进一步补充Fgf10,胃泌素,Wnt3a和更高浓度的R-spondin 1以获得生产性 - 文化。相比之下,到目前为止,很少知道在发育期间排列前腺上皮细胞的胎儿细胞。到目前为止,它们的性质以及其离体的潜在生长特性未明确。基于以前的研究,确定胎儿小肠(Mustata等人,2013年)中存在的细胞,我们报告了作为球体的小鼠胎儿胃祖细胞的培养(Fernandez Vallone et al。 。,2016)。可以在2009年由佐藤等人先前报道的培养基中重复胃祖细胞以生长小肠成体干细胞,与成人型胃干细胞相反,它们不需要额外的生长因子补充(Fgf10,Wnt3a或胃泌素)。 ...

Relative Stiffness Measurements of Cell-embedded Hydrogels by Shear Rheology in vitro
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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