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Insulin from bovine pancreas

来自牛胰腺的胰岛素

Company: Sigma-Aldrich
Catalog#: I6634
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Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma
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Date:
2017-09-05
[Abstract]  Most epithelial tumors have been shown to contain cancer stem cells that are potentially the driving force in tumor progression and metastasis (Kreso and Dick, 2014; Nassar and Blanpain, 2016). To study these cells in depth, cell isolation strategies relying on cell surface markers or fluorescent reporters are essential, and the isolation strategies must preserve their viability. The ability to isolate different populations of cells from the bulk of the tumor will continue to deepen our understanding of the biology of cancer stem cells. Here, we report the strategy combining mechanical tumor dissociation, enzymatic treatment and flow cytometry to isolate a pure population of epithelial cancer stem cells from their native microenvironment. This technique can be useful to further ... [摘要]  大多数上皮肿瘤已经显示含有可能是肿瘤进展和转移的驱动力的癌症干细胞(Kreso和Dick,2014; Nassar和Blanpain,2016)。 为了深入研究这些细胞,依赖于细胞表面标志物或荧光报告基因的细胞分离策略是必不可少的,分离策略必须保持其活力。 从大部分肿瘤中分离不同细胞群的能力将继续加深我们对癌症干细胞生物学的认识。 在这里,我们报告了结合机械肿瘤解离,酶处理和流式细胞术的策略,从其天然微环境中分离出纯种群的上皮癌干细胞。 该技术可用于进一步功能性地分析癌症干细胞(RNA测序和表观遗传学分析),在培养物中培养它们或在移植测定中直接使用它们。
【背景】肿瘤复发和转移是大多数与癌症有关的死亡的主要原因。恶性肿瘤可能由干细胞群体启动和维持(Nassar和Blanpain,2016; Bonnet和Dick,1997),这些细胞是预防复发的重要治疗靶点(Baumann et al。,2008)。研究表明,鳞状细胞癌由肿瘤干细胞亚群维持,其抗药性,并通过进行自我更新和分化(如正常干细胞)引发肿瘤复发,产生增殖祖细胞,其分化形成肿瘤大部分(Locke et al。,2005; Prince et al。,2007; Malanchi et al。,2008; de Sousa e Melo et ...

Melanoma Stem Cell Sphere Formation Assay
Author:
Date:
2017-04-20
[Abstract]  Self-renewal is the ability of cells to replicate themselves at every cell cycle. Throughout self-renewal in normal tissue homeostasis, stem cell number is maintained constant throughout life. Cancer stem cells (CSCs) share this ability with normal tissue stem cells and the sphere formation assay (SFA) is the gold standard assay to assess stem cells (or cancer stem cells) self-renewal potential in vitro. When single cells are plated at low density in stem cell culture medium, only the cells endowed with self-renewal are able to grow in tridimensional clusters usually named spheres. In the recent years, SFA has also been used also to test the effect of several drugs, chemical and natural compounds or microenviromental components on stem cells self-renewal capacity. Here we will ... [摘要]  自我更新是细胞在每个细胞周期复制自身的能力。在正常组织体内平衡的自我更新过程中,干细胞数量在整个生命中保持不变。癌症干细胞(CSCs)与正常组织干细胞共享这种能力,球形成测定(SFA)是评估干细胞(或癌症干细胞)体外自我更新潜力的金标准测定方法。 。当单细胞在干细胞培养基中以低密度铺板时,仅具有自我更新的细胞能够在通常称为球体的三维簇中生长。近年来,SFA也用于测试几种药物,化学和天然化合物或微环境成分对干细胞自我更新能力的影响。在这里,我们将说明一个详细的方案来评估人类黑色素瘤干细胞的自我更新,作为黑色球体生长。

癌症干细胞(CSCs)首先在急性骨髓瘤白血病(Lapidot et al。,1994)中发现,然后在许多实体瘤中鉴定出包括黑素瘤。 CSCs被定义为具有自我更新和肿瘤起始能力的细胞,能够在体内再生整个肿瘤异质性。可以使用基于表型特征或生物学特性的不同方法从肿瘤块中分离CSCs,然后在体外(自我更新)和体内 (致瘤潜力)。使用细胞表面标志物的组合分离黑素瘤CSCs(Fang等人,2005; Monzani等人,2007; Schatton等人, ,2008; Boiko等人,2010; Boonyaratanakornkit等人,2010)或通过特定干细胞培养基中的培养(Perego等人, ,2010; Santini ...

Dissociated Retinal Cell Culture
Author:
Date:
2014-01-20
[Abstract]  The retina is a relatively simple and accessible part of the central nervous system, making it a powerful model to study cell fate specification mechanisms. Multipotent retinal progenitor cells (RPCs) give rise to seven major classes of retinal cell types. Mechanisms regulating cell fate choice in the retina depend on both cell intrinsic and environmental factors, but their relative contribution to specific cell fate decisions remains unclear. Dissociated retinal cell cultures provide a great assay to study this problem. RPCs are cultured in serum-free and extract-free medium, providing the investigator with a control over the environment to address questions related to the effects of a particular molecule on the development of retinal neurons. In addition, dissociated cell cultures can ... [摘要]  视网膜是中枢神经系统相对简单易用的部分,是研究细胞命运规范机制的有力模型。多能视网膜祖细胞(RPC)产生七大类视网膜细胞类型。调节视网膜细胞命运选择的机制取决于细胞内在和环境因素,但它们对特定细胞命运决定的相对贡献仍不清楚。分离的视网膜细胞培养物提供了一个很好的分析来研究这个问题。 RPC在无血清和无提取物培养基中培养,为研究者提供对环境的控制,以解决与特定分子对视网膜神经元发育的影响有关的问题。此外,分离的细胞培养物可用于通过从其正常环境中分离RPC来研究细胞内在机制的重要性(Cayouette等人,2003; Jensen和Raff,1997)。下述方法适用于RPC的克隆密度培养。在这样的培养物中,RPC彼此分离并且从后神经元分离。它们分化和分化成不同的视网膜细胞类型以形成小菌落或“克隆”。在最近的一项研究中,我们发现这些克隆与细胞数量和细胞类型组成方面在视网膜原位发生的克隆是无法区分的,这表明细胞内机制在视网膜发育中起关键作用(Cayouette et al。 ,2003)。

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