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PE anti-human CD63 Antibody

Company: BioLegend
Catalog#: 353003
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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 ...

Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures
Author:
Date:
2017-09-05
[Abstract]  Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy is regulated provides an opportunity to explore druggable targets and subsequently develop treatment strategies for these diseases. To identify novel autophagy regulators, we developed an image-based phenotypic RNAi screening approach using autophagic marker proteins at endogenous levels (Jung et al., 2017). In contrast to previously performed autophagy screens, this approach does not use overexpressed, tagged autophagy marker proteins ... [摘要]  自噬是一种循环途径,其中细胞内货物包括蛋白质聚集体和细菌被自噬吞噬,随后与溶酶体融合后降解。这种过程的失调涉及几种人类疾病,如癌症或神经退行性疾病。因此,提高我们对自噬如何监管的理解提供了探索可药用靶标的机会,并随后制定了这些疾病的治疗策略。为了鉴定新的自噬调节因子,我们开发了一种基于图像的表型RNAi筛选方法,在内源水平上使用自噬标记蛋白(Jung et al。,2017)。与以前进行的自噬屏幕相比,该方法不使用过表达标记的自噬标记蛋白,而是在内源水平检测自噬结构。此外,我们同时监测自噬的早期和晚期阶段,而其他筛选仅使用单个自噬体标志物大多为GFP-LC3B。在这里,我们详细描述了这种多重筛选方案,并讨论了如何建立基于图像的siRNA筛选的一般考虑。
【背景】自噬是一种细胞内质量和数量控制途径,通过这种途径,多种细胞溶质材料如病原体,细胞器或其部分,蛋白质和其他大分子被双重膜结构所吞噬,造成自噬体,并在自噬体与溶酶体融合后进行大量溶酶体降解。自体吞噬体的形成及其对自体溶酶体的成熟是一个高度规范的过程。在酵母中最初鉴定的AuTophaGy相关(ATG)基因是泛蛋白样蛋白Atg8,其通过与位于受体自噬体中的磷脂磷脂酰乙醇胺(PE)的可逆缀合以高度局部化的方式发挥其功能。人类细胞含有六个ATG8家族成员,可以分为两个亚科:i)微管相关蛋白1A / ...

Glioma Induction by Intracerebral Retrovirus Injection
Author:
Date:
2017-07-20
[Abstract]  Glioblastoma (GBM) is the most common primary brain cancer in adults and has a poor prognosis. It is characterized by a high degree of cellular infiltration that leads to tumor recurrence, atypical hyperplasia, necrosis, and angiogenesis. Despite aggressive treatment modalities, current therapies are ineffective for GBM. Mouse GBM models not only provide a better understanding in the mechanisms of gliomagenesis, but also facilitate the drug discovery for treating this deadly cancer. A retroviral vector system that expresses PDGFBB (Platelet-derived growth factor BB) and inactivates PTEN (Phosphatase and tensin homolog) and P53 tumor suppressors provides a rapid and efficient induction of glioma in mice with full penetrance. In this protocol, we describe a simple and practical method for ... [摘要]  成胶质细胞瘤(GBM)是成人中最常见的原发性脑癌,预后差。其特征在于高度的细胞浸润,导致肿瘤复发,非典型增生,坏死和血管生成。尽管采取积极的治疗方式,目前的疗法对GBM无效。小鼠GBM模型不仅提供了对胶质瘤发生机制的更好理解,而且有助于药物发现治疗这种致命的癌症。表达PDGFBB(血小板衍生生长因子BB)和灭活PTEN(磷酸酶和张力蛋白同源物)和P53肿瘤抑制因子的逆转录病毒载体系统提供了具有全面外显子的小鼠中胶质瘤的快速和有效的诱导。在该方案中,我们描述了一种简单实用的方法,用于在鼠脑中通过逆转录病毒注射诱导GBM形成。该系统对神经胶质瘤的诱导进行空间和时间控制,并允许用生物发光报告物评估治疗效果。
【背景】胶质母细胞瘤(GBM)是最具侵袭性的恶性脑肿瘤,不幸的是几乎总是致命的。尽管多种治疗方式有进展,但是尚未开发出治疗GBM的有效治疗方法。这种疾病的机制仍然知之甚少。动物模型一直是定义GBM发病机制和基因或药物治疗试验的非常重要的工具。已经开发了几种小鼠模型,其目的是产生尽可能接近模拟人类疾病的疾病,并表现出类似的分子,遗传和组织学特征。突出使用的模型是异种移植(Hingtgen等人,2008)模型,其中人类肿瘤细胞系可以在脑中原位移植以及在免疫受损的小鼠的皮下区域异位移植,提供了具有大肿瘤块在短时间内。遗传工程小鼠模型(GEMM)具有特异性增益的致癌活性或丧失肿瘤抑制通路,其类似于GBM中最频繁失调的通路中的扰动,导致啮齿类动物形成胶质瘤。 ...

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