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Ficoll-Paque PLUS

Ficoll-Paque PLUS

Company: GE Healthcare
Catalog#: 17-1440
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Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)
Author:
Date:
2016-11-20
[Abstract]  The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy. [摘要]  本协议的目的是描述如何使用三种不同的技术:流式细胞术,定量PCR和共聚焦显微镜来测量和量化人类单核细胞衍生树突细胞(MDDCs)中内在的HIV-1颗粒和葡聚糖分子的量。

[背景] 开发此协议是为了评估在人单核细胞衍生的树突细胞中肌动蛋白成核破坏时HIV-1内化的变化。在shRNA筛选后,识别对于HIV-1从树突状细胞转移到T细胞重要的基因,我们观察到肌动蛋白成核的中断导致从富含肌动蛋白的树突到水泡的转变,由于过量的肌动球蛋白收缩。结果,观察到HIV-1转移的减少和由于水泡收缩驱动的巨噬细胞增多引起的HIV-1内化的增加。我们的结论是,肌动蛋白成核和稳定的效应器是维持艾滋病毒1对肌动蛋白丰富的树突和限制其内吞作用,有效转移到T淋巴细胞的关键(Menager和Littman,2016)。

Isolation and Primary Culture of Various Cell Types from Whole Human Endometrial Biopsies
Author:
Date:
2016-11-20
[Abstract]  The isolation and primary culture of cells from human endometrial biopsies provides valuable experimental material for reproductive and gynaecological research. Whole endometrial biopsies are collected from consenting women and digested with collagenase and DNase I to dissociate cells from the extracellular matrix. Cell populations are then isolated through culturing, filtering and magnetic separation using cell-surface antigen markers. Here we provide a comprehensive protocol on how to isolate and culture individual cell types from whole endometrial tissues for use in in vitro experiments. [摘要]  从人子宫内膜活组织检查中分离和原代培养细胞为生殖和妇科研究提供了有价值的实验材料。从同意的妇女收集完整的子宫内膜活检组织并用胶原酶和DNA酶I消化以从细胞外基质中分离细胞。然后通过使用细胞表面抗原标记物的培养,过滤和磁性分离来分离细胞群。在这里,我们提供了如何从整个子宫内膜组织分离和培养个体细胞类型以用于体外实验的全面方案。

[背景] 人子宫内膜是子宫内层最粘的层。其由柱状上皮和基础基质层组成,其经历循环再生,生长和响应于循环激素的转化。子宫内膜衬里分化为腺分泌表型为囊胚植入和成功怀孕提供了一个好客的环境。在没有怀孕这个层脱落,导致月经。从人子宫内膜活组织检查中分离和培养细胞允许在体外功能评估和与患者结果相关的细胞特征的研究。子宫内膜细胞的分离和培养是研究妇科和产科医学的许多方面的非常宝贵的研究模型,包括不孕,植入失败,复发性流产和月经紊乱。全人子宫内膜活检包含人子宫内膜基质细胞(HESCs),腔和子宫内膜上皮细胞(HEEC),红细胞和免疫细胞的混合群体。 ...

Efficient Isolation of Influenza Specific CTLs
Author:
Date:
2016-06-20
[Abstract]  Human antigen-specific CD8+ T-cell clones are valuable tools for dissecting CD8+ T-cell responses against antigens derived from infectious agents, cancer and self antigens. Here we describe a protocol for isolating human antigen-specific CD8+ T cells. This protocol uses surface capture of IFNγ to identify antigen responsive cells that are then cloned by single-cell sorting. Here we use CD8+ T-cell responses to influenza matrix protein (MP) as an example, but this approach can be applied to any antigen specificity. [摘要]  人抗原特异性CD8 + T细胞克隆是用于解剖来自感染剂,癌症和自身抗原的抗原的CD8 + T细胞应答的有价值的工具。 在这里我们描述了用于分离人抗原特异性CD8 + T细胞的方案。 该协议使用IFNγ的表面捕获来鉴定抗原反应性细胞,然后通过单细胞分选克隆。 这里我们使用CD8 + T细胞对流感基质蛋白(MP)的反应作为例子,但这种方法可以应用于任何抗原特异性。

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