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RPMI1640 medium

RPMI MEDIUM 1640(CE)

Company: Thermo Fisher Scientific
Catalog#: 21875034
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A Flow-assay for Farnesol Removal from Adherent Candida albicans Cultures
Author:
Date:
2017-10-05
[Abstract]  Here, we describe a protocol for a continuous flow system for C. albicans cultures growing adherent to a plastic surface. The protocol was adapted from a previous method established to simulate blood flow on endothelial cells (Wilson and Hube, 2010). The adapted protocol was used by us for the removal of molecules in C. albicans supernatants, especially farnesol, which accumulate over the time course of incubation and cannot be specifically depleted. The system used, however, allows various applications including the simulation of physiological flow conditions. Several example applications are given on the manufacturer’s website (https://ibidi.com/perfusion-system/112-ibidi-pump-system.html ... [摘要]  在这里,我们描述了用于连续流系统的协议。 白色念珠菌生长粘附在塑料表面上的培养物。 该方案根据建立以模拟内皮细胞上的血液流动的先前方法改编(Wilson和Hube,2010)。 我们使用适应方案去除C中的分子。 白色念珠菌上清液,特别是法呢醇,其在孵育的时间过程中积累并且不能被特异性地耗尽。 然而,所使用的系统允许各种应用,包括模拟生理流动条件。 制造商网站上给出了几个示例应用程序(https://ibidi.com/perfusion-system/112-ibidi-pump-system.html).
【背景】法尼醇是人类致病真菌白色念珠菌中的酵母 - 菌丝转移(Hornby等人,2001)的有效抑制剂,并且还促进了酵母生长的逆转预制长丝(Lindsay等人,2012)。群体感知分子(QSM)快速积聚在白念珠菌EED1缺失株的上清液中,并促进突变体(Polke等人)的反向形态发生和菌丝维持缺陷>,2017)。由于我们无法阻止法呢醇的合成(Polke等人,2017),我们使用了ibidi ®泵系统,通过单向去除上清液中累积的QSMs流。流动应用以及在孵育期间的恒定的培养基更换在C中显着延长了丝状化。白色念珠菌eed1Δ突变体。这表明QSM积累的成功去除,并且提供了在C中的菌丝维持和法呢醇信号之间的直接联系。白色假丝酵母。用于此协议(ibidi ...

In vitro Demonstration and Quantification of Neutrophil Extracellular Trap Formation
Author:
Date:
2017-07-05
[Abstract]  In the recent decade, neutrophil extracellular traps (NETs) have been identified and confirmed as a new anti-microbial weapon of neutrophils. In this protocol, we describe easy methods to demonstrate NET formation by immunofluorescence staining of extracellular chromatin fiber with anti-DNA/Histone H1 antibody and quantification of NETs by using a non-cell-permeable DNA specific dye Sytox orange. [摘要]  近十年来,嗜中性粒细胞胞外捕获物(NETs)已被鉴定并确认为一种新的抗微生物中性粒细胞武器。 在该方案中,我们描述了通过使用抗DNA /组蛋白H1抗体的细胞外染色质纤维的免疫荧光染色和通过使用非细胞可渗透的DNA特异性染料Sytox orange来定量NETs的简便方法来证明NET形成。
【背景】嗜中性粒细胞构成循环白细胞中最大的进化保守部分。他们通过各种机制建立了针对病原体的第一道防线,包括形成嗜中性粒细胞胞外捕获物(NETs)。在此过程中,活化的嗜中性粒细胞从细胞核中排出染色质纤维。入侵的病原体然后被捕获在染色质网络内并被高度浓缩的NET缠结的抗微生物蛋白如髓过氧化物酶(MPO)和弹性蛋白酶(Brinkmann等人,2004)杀死。然而,NETs是一把双刃剑;来自过度活化的嗜中性粒细胞的无限制的NET形成也可能导致严重的组织损伤,例如通过NET的组蛋白组分的细胞毒性作用(Saffarzadeh等人,2012)。中性粒细胞过度活化并具有增强的形成NETs的能力的病理状况的一个例子是系统性红斑狼疮。狼疮患者血清中针对双链DNA及其它组分的抗体水平升高(Knight and Kaplan,2012; Yu and ...

Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC)
Author:
Date:
2017-05-05
[Abstract]  Mesenchymal stem/stromal cells (MSC) are adult stem cells which have been shown to improve survival, enhance bacterial clearance and alleviate inflammation in pre-clinical models of acute respiratory distress syndrome (ARDS) and sepsis. These diseases are characterised by uncontrolled inflammation often underpinned by bacterial infection. The mechanisms of MSC immunomodulatory effects are not fully understood yet. We sought to investigate MSC cell contact-dependent communication with alveolar macrophages (AM), professional phagocytes which play an important role in the lung inflammatory responses and anti-bacterial defence. With the use of a basic direct co-culture system, confocal microscopy and flow cytometry we visualised and effectively quantified MSC mitochondrial transfer to AM ... [摘要]  间充质干/基质细胞(MSC)是成年干细胞,已被证明可以改善急性呼吸窘迫综合征(ARDS)和败血症临床前模型中的存活,增强细菌清除和减轻炎症。这些疾病的特征在于通常受细菌感染支持的不受控制的炎症。 MSC免疫调节作用的机制尚未完全了解。我们试图研究与肺泡巨噬细胞(AM),在肺炎症反应和抗菌防御中起重要作用的专业吞噬细胞的MSC细胞接触依赖性通信。通过使用基本的直接共培养系统,共聚焦显微镜和流式细胞术,我们通过隧道纳米管(TNT)显现并有效量化了MSC线粒体转移到AM。为了模拟人类AM,在粒细胞巨噬细胞集落刺激因子(GM-CSF)存在下,从人供体血液中分离原代单核细胞并分化成巨噬细胞(单核细胞衍生的巨噬细胞,MDM),从而允许适应AM样表型(de Almeida等人,2000; Guilliams等人,2013)。人骨髓衍生的MSC用线粒体特异性荧光染色标记,广泛洗涤,以1:20(MSC / ...

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