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Falcon® 96-well Clear Flat Bottom TC-treated Culture Microplate

Falcon® 96孔透明平底TC处理培养微孔板

Company: Corning
Catalog#: 353072
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Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage
Author:
Date:
2017-08-20
[Abstract]  Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria producing colibactin following a short infection of cultured mammalian cells with pks+ E. coli. [摘要]  具有pks基因组岛的大肠杆菌菌株合成基因毒素大肠杆菌素。 将真核细胞暴露于产生大肠杆菌的大肠杆菌诱导DNA损伤,最终导致细胞周期停滞,衰老和死亡。 在这里,我们描述了一种简单的方法来证明在用pks +大肠杆菌培养的哺乳动物细胞的短时间感染后,产生大肠杆菌素的细菌的遗传毒性。
【背景】大肠杆菌素是在大肠杆菌的肠外致病,共生和益生菌菌株中发现的基因毒素(Nougayrede等,2006)。大肠杆菌素也由其他肠杆菌科产生,包括肺炎克雷伯杆菌,产气肠杆菌和柠檬酸杆菌(Putze等人,2009)。大肠杆菌素是通过由多酮酶和非核糖体肽合酶(PKS和NRPS),定制和成熟酶以及外排泵组成的多酶机械合成的聚酮化合物/非核糖体肽杂化化合物(综述:Taieb等人。,2016)。该合成机制编码在52kb的基因组,即'pks'岛上。 Colibactin在感染pks +细菌的真核细胞中诱导DNA损伤。由大肠杆菌素诱导的遗传毒性作用需要活的pks +细菌与真核细胞的直接接触。事实上,杀死的细菌或细菌上清液或裂解物没有观察到遗传毒性作用。因此,为了证明产生大肠杆菌的大肠杆菌的基因毒性,培养的哺乳动物细胞(例如HeLa细胞)在4小时内用活的pks ...

In vitro Assay to Assess Efficacy of Potential Antiviral Compounds against Enterovirus D68
Author:
Date:
2017-03-20
[Abstract]  In 2014 enterovirus D68 (EV-D68) caused the largest outbreak in the United States since the discovery of the virus. Distinct from before, the 2014 infections were associated with more severe respiratory disease and occasional neurological complications. So far, there are no available vaccines or antivirals for the prophylaxis or treatment of EV-D68 infections. In order to evaluate the antiviral activity of potential inhibitors of EV-D68 replication, a cell-based cytopathic effect (CPE) reduction assay was developed (Sun et al., 2015). [摘要]  2014年肠病毒D68(EV-D68)自发现病毒以来,在美国引起了最大的爆发。与之前不同的是,2014年感染与更严重的呼吸道疾病和偶尔的神经系统并发症有关。到目前为止,还没有可用的疫苗或抗病毒药物用于预防或治疗EV-D68感染。为了评估EV-D68复制潜在抑制剂的抗病毒活性,开发了基于细胞的细胞病变效应(CPE)降低测定法(Sun等,2015)。

背景 作为新出现的病原体,以前很少报道靶向EV-D68的抗病毒化合物。迫切需要建立和开发抗病毒方法来对抗潜在的EV-D68流行病。在这里,我们报告一个详细的方案,可用于识别选择性抗EV-D68化合物。为了筛选和鉴定潜在的抗病毒化合物,基于MTS的CPE降解测定法是可重复使用的,易于使用和省时的方法,被广泛使用。该方法依赖于化合物抑制EV-D68诱导的CPE降低的能力。当化合物主动抑制EV-D68的复制时,病毒诱导的CPE将被减少或不存在。因此,代谢活性细胞能够将黄色四唑盐底物(MTS / PMS)转化为棕色甲an制品。当化合物不具有抗病毒作用时,宿主细胞死于病毒诱导的CPE,其将缺乏代谢活性,并且黄色底物保持未代谢。比色转化是定量的,因为可以从该测定产生的数据计算EC 50。

Isolation of Murine Adipose Tissue-derived Mesenchymal Stromal Cells (mASCs) and the Analysis of Their Proliferation in vitro
Author:
Date:
2015-11-05
[Abstract]  Mesenchymal stromal cells (MSCs) are non-hematopoietic, perivascular cells which support hematopoiesis and are thought to participate in tissue repair in vivo. MSCs can be isolated from various tissues and organs and are defined in vitro as plastic adherent cells expressing CD73, CD90, CD105 (human MSCs) or CD29, CD44, sca-1 (murine MSCs) which can differentiate into osteoblasts, adipocytes, chondroblasts and myocytes. MSCs possess potent immunomodulatory and trophic capacities in vitro and in vivo and have thus emerged as a promising treatment of inflammatory/autoimmune diseases. The use of MSCs for human disease relies on the injection of a large number of cells and much effort has been focused on acquiring MSCs with high proliferative capacity. Thus, ... [摘要]  间充质基质细胞(MSC)是非造血的血管周围细胞,其支持造血并且被认为在体内参与组织修复。 MSC可以从各种组织和器官中分离并在体外定义为表达CD73,CD90,CD105(人MSC)或CD29,CD44,sca-1(鼠MSC)的塑性贴壁细胞,其可以分化成骨细胞,脂肪细胞,软骨细胞和肌细胞。 MSC在体外具有强的免疫调节和营养能力,并且在体内具有强的免疫调节能力,因此已经成为炎症/自身免疫疾病的有希望的治疗。 MSC对人类疾病的使用依赖于大量细胞的注射,并且许多努力已经集中在获得具有高增殖能力的MSC。因此,建立简单和准确的测定MSC增殖的协议对于基础研究和应用研究都是重要的。目前的方案提供了关于如何使用xCELLigence系统和CellTiter-Blue试剂(Carrillo-Galvez等人,)分离和测量源自腹股沟和/或腹内脂肪组织(mASCs)的鼠MSC的增殖的细节>,2015; Anderson ,,2013)。下面描述的方案也可以容易地翻译成人MSC。

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