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Falcon® 60 mm TC-treated Cell Culture Dish

Falcon® 60mmTC细胞培养皿

Company: Corning
Catalog#: 353002
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Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma
Author:
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 ...

Antisense Oligonucleotide-mediated Knockdown in Mammary Tumor Organoids
Author:
Date:
2017-08-20
[Abstract]  Primary mammary tumor organoids grown in 3D are an excellent system to study tumor biology. They resemble the organization and physiology of native epithelia more closely than cancer cell lines grown in 2D, and additionally model interactions with the ECM (Boj et al., 2015; Clevers, 2016; Shamir and Ewald, 2014). Mammary tumor organoids are therefore a promising model system to identify and characterize novel drivers of breast cancer that would be unlikely to be identified using 2D cell lines. Antisense oligonucleotides can be used to efficiently and specifically knockdown target genes in the cell (Bennett et al., 2017). They can be taken up freely by organoids without the need for a transfection agent, making them a convenient tool for routine lab studies and screens. [摘要]  在3D生长的原发性乳腺肿瘤组织是研究肿瘤生物学的优秀系统。 它们类似于天然上皮的组织和生理学,比2D生长的癌细胞系更为紧密,另外还与ECM的模型相互作用(Boj et al。,2015; Clevers,2016; Shamir and Ewald,2014)。 因此,乳腺肿瘤组织因子是一种有希望的模型系统,用于识别和表征不可能使用2D细胞系识别的乳腺癌的新型驱动因素。 反义寡核苷酸可用于有效和特异地敲低细胞中的靶基因(Bennett等,2017)。 它们可以被有机物自由摄取,而不需要转染剂,使其成为常规实验室研究和筛选的便捷工具。
【背景】乳腺癌是全世界妇女中最常见的恶性肿瘤,是妇女癌症死亡率的第二大原因(Siegel等,2017)。为了改善现有的治疗方案,确定和调查具有预防乳腺癌进展潜力的新分子靶标至关重要。我们应用RNA-seq来产生与正常乳腺上皮细胞相比在原发性乳腺肿瘤中失调的长非编码RNA(lncRNA)的综合目录,并将30个先前未表征的lncRNA作为乳腺肿瘤相关RNA(MaTARs)进行优先排序。为了功能评估MaTARs作为肿瘤进展的关键驱动因素,我们对3D乳腺肿瘤组织中的所有30个MaTARs进行了反义寡核苷酸(ASO)介导的敲低分析(Diermeier等,2016)。
   ASO是短(20-mers),含有硫代磷酸酯修饰的核苷酸的单链DNA分子以及2'-ribose(5-10-5 ...

Preparation of Mosquito Salivary Gland Extract and Intradermal Inoculation of Mice
Author:
Date:
2017-07-20
[Abstract]  Mosquito-transmitted pathogens are among the leading causes of severe disease and death in humans. Components within the saliva of mosquito vectors facilitate blood feeding, modulate host responses, and allow efficient transmission of pathogens, such as Dengue, Zika, yellow fever, West Nile, Japanese encephalitis, and chikungunya viruses, as well as Plasmodium parasites, among others. Here, we describe standardized methods to assess the impact of mosquito-derived factors on immune responses and pathogenesis in mouse models of infection. This protocol includes the generation of mosquito salivary gland extracts and intradermal inoculation of mouse ears. Ultimately, the information obtained from using these techniques can help reveal fundamental mechanisms of interaction between ... [摘要]  蚊子传播的病原体是人类严重疾病和死亡的主要原因之一。蚊子唾液中的组分促进血液供体,调节宿主反应,并允许有效传播病原体,如登革热,紫草,黄热病,西尼罗河,日本脑炎和基孔肯雅病毒,以及疟原虫寄生虫等。在这里,我们描述了评估蚊子衍生因子对小鼠感染模型中免疫反应和发病机制的影响的标准化方法。该方案包括产生蚊子唾液腺提取物和皮内接种小鼠耳朵。最终,使用这些技术获得的信息可以帮助揭示病原体,蚊子和哺乳动物宿主之间的相互作用的基本机制。此外,该协议还可以帮助建立疫苗或治疗剂的临床前检测改进的感染挑战模型,以考虑通过蚊子传播的自然途径。
【背景】在探测血液的同时,蚊子接种有助于喂养的唾液,但如果蚊子曾经感染过感染个体,也可能含有病原体。蚊子唾液在建立感染,促进传播,调节免疫应答和加剧西尼罗河病毒发病过程中发挥重要作用(Schneider等人,2006; Styer等人)。 ,2011),登革热病毒(Cox等人,2012; Conway等人,2014; McCracken等人,2014; Schmid ,2016),基孔肯雅病毒(Agarwal等人,2016),Semliki Forest病毒(Pingen等人,2016) ,裂谷热病毒(Le Coupanec等人,2013)和疟原虫寄生虫(Schneider等人,2011)感染。许多重要问题仍然存在,需要改进动物模型。
 而通过感染的蚊子接种最好地模仿自然传播,接种剂量的高度变异性和有限的昆虫设施的可用性导致这种程序的有限使用。此外,使用感染的蚊子时,不能控制唾液的含量和蚊子组分的存在或不存在。作为替代方案,未感染的雌性蚊子的“斑点喂养”,然后通过针对病原体进行真皮内接种,模拟了唾液在小鼠皮肤中的自然沉积并递送了一定剂量的病原体。 ...

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