| Isolation and Primary Cell Culture of Mouse Dorsal Root Ganglion Neurons
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Author:
Date:
2016-04-05
[Abstract] We here provide a detailed protocol for the isolation and culture of primary mouse sensory neurons. The cell bodies of sensory afferent pseudounipolar neurons are located in dorsal root ganglia (DRGs) along the vertebral column. Dissected mouse DRGs can be dissociated into single cells by enzymatic digestion to obtain primary cultures of mouse sensory neurons as performed in the studies reported by Khaminets et al. (2015).
[摘要] 我们在这里提供了详细的协议，用于隔离和培养的主要小鼠感觉神经元。 感觉传入假性极化神经元的细胞体位于沿着脊柱的背根神经节（DRG）中。 解离的小鼠DRG可以通过酶消化解离成单个细胞，以获得小鼠感觉神经元的原代培养物，如Khaminets等人（2015）报道的研究中所进行的。
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| Ex vivo Human Natural Killer (NK) Cell Stimulation and Intracellular IFNγ and CD107a Cytokine Staining
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Author:
Date:
2015-06-20
[Abstract] Natural killer (NK) cells comprise 5–20% of peripheral blood mononuclear cells (PBMC) in humans. In addition to their fundamental roles in the defense against viral infections and tumor surveillance, NK cells help shape adaptive immune responses through their production of cytokines. NK cells are traditionally identified as CD3neg, CD14neg, CD19neg lymphocytes expressing CD56. Using a combination of markers that includes CD56 and CD7 greatly increases the ability to define the phenotype and function of NK cell subsets. Two key markers of NK cell function are ...
[摘要] 自然杀伤（NK）细胞在人中包含5-20％的外周血单核细胞（PBMC）。 除了它们在防御病毒感染和肿瘤监测中的基本作用，NK细胞通过其细胞因子的产生帮助形成适应性免疫应答。 NK细胞传统上被鉴定为表达CD56的CD3阴性，CD14阳性，CD19阴性淋巴细胞。 使用包括CD56和CD7的标记物的组合极大地增加了定义NK细胞亚群的表型和功能的能力。 NK细胞功能的两个关键标记是IFNγ的产生和通过CD107a的表达测量的细胞毒性颗粒的释放。 在这里我们描述了一种方法来评估在靶细胞或细胞因子刺激后NK细胞中的IFNγ和CD107a表达。 该方法可用于评估来自广泛的研究参与者的外周血单核细胞中NK细胞的一般功能能力。
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| Macrophage Inflammatory Assay
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Author:
Date:
2014-07-20
[Abstract] Macrophages represent a widely distributed and functionally diverse population of innate myeloid cells involved in inflammatory response to pathogens, tissue homeostasis and tissue repair (Murray and Wynn, 2011). Macrophages can be broadly grouped into two subpopulations with opposing activites: M1 or pro-inflammatory macrophages that promote T-helper type 1 (Th1) cell immunity and tissue damage, and M2 or anti-inflammatory/alternatively activated macrophages implicated in Th2 response and resolution of inflammation. Here we describe a rapid assay we used previously to monitor changes in ...
[摘要] 巨噬细胞代表广泛分布的和功能不同的先天骨髓细胞群，参与对病原体的炎症反应，组织内稳态和组织修复（Murray和Wynn，2011）。巨噬细胞可以大致分为具有相反活性的两个亚群：M1或促炎性巨噬细胞，其促进T辅助1型（Th1）细胞免疫和组织损伤，以及M2或抗炎或交替激活的巨噬细胞涉及Th2反应和分辨率的炎症。在这里我们描述了一种快速测定，我们以前用于监测由脂多糖（LPS）激活的巨噬细胞在响应于由成体干细胞产生的治疗性旁分泌因子的促炎和抗炎细胞因子产生中的变化（Bartosh等，/em>，2010; Ylostalo等人，2012; Bartosh 等人，2013）。该测定可以适当地适应于测试巨噬细胞对其它试剂的响应，在本文中将称为"测试试剂"或"测试化合物"。在该方案中，小鼠巨噬细胞细胞系J774A.1被扩增作为陪替氏培养皿上的粘附单层，允许容易地收获细胞而没有可以损伤细胞的酶或细胞刮擦器。然后用LPS悬浮刺激大孔，并接种到含有试验试剂的12孔细胞培养板中。 16-18小时后，收集由巨噬细胞调节的培养基，并用酶联免疫吸附测定（ELISA）测定培养基中的细胞因子谱。我们常规测量促炎细胞因子TNF-α和抗炎细胞因子白细胞介素-10（IL-10）的水平。
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