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Company: Thermo Fisher Scientific
Catalog#: 21051024
Other protocol()

Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma
[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 ...

Isolation and Analysis of Stromal Cell Populations from Mouse Lymph Nodes
[Abstract]  Our protocol describes a simple procedure for isolating stromal cells from lymph nodes (LN). LN are disrupted then enzymatically digested with collagenase and dispase to produce a single cell suspension that can be stained with fluorescently labelled antibodies and analysed by flow cytometry. This protocol will enable identification of fibroblastic reticular cells (FRC), lymphatic endothelial cells (LEC), blood endothelial cells (BEC) as PNAd+ BEC that form LN high endothelial venules (HEV). This method can be applied to examine LN stromal cell responses during inflammatory events induced by infections or immunologic adjuvants and to subset most leukocytes found in LN. [摘要]  我们的方案描述了从淋巴结(LN)分离基质细胞的简单过程。 LN被破坏,然后用胶原酶和分散酶酶消化以产生可以用荧光标记的抗体染色的单细胞悬浮液并通过流式细胞术分析。 该方案能够识别形成LN高内皮小静脉(HEV)的PNAd + BEC的成纤维细胞网状细胞(FRC),淋巴内皮细胞(LEC),血液内皮细胞(BEC)。 该方法可以用于检测由感染或免疫佐剂诱导的炎症事件期间的LN基质细胞反应,并且在LN中发现大多数白细胞。

In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor
[Abstract]  Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger et al., 2014). This protocol is designed for treatment with Kyn or FICZ in mouse embryonic fibroblasts (MEFs) or primary peripheral monocytes. [摘要]  通过内源性配体活化芳基烃受体(AHR)已涉及多种生理过程,如细胞周期调控,细胞分化和免疫应答。据报道,色氨酸代谢物,如犬尿胆碱(Kyn)和6-甲基吲哚(3,2-b)咔唑(FICZ)是AHR的内源性配体(Stockinger等人,2014)。该方案设计用于在小鼠胚胎成纤维细胞(MEF)或初级周边单核细胞中用Kyn或FICZ进行治疗。

背景 色氨酸代谢物如Kyn和FICZ是生理条件下AHR的内源性配体。 Kyn由色氨酸-2,3-双加氧酶(TDO)和/或吲哚胺-2,3-双加氧酶1和2(IDO1 / 2)产生,并有助于抑制抗肿瘤反应和恶性进展(Stockinger等人,2014)。 ...