Isolation and Culture of Human Endometrial Epithelial Cells and Stromal Fibroblasts
|
Author:
Date:
2015-10-20
[Abstract] Purification and culture of endometrial epithelial cells (eEC) and stromal fibroblasts (eSF) from endometrial biopsies allows for downstream cell-specific in vitro studies. The utility of this protocol is the ease with which cells are purified without contamination from unwanted cell types, and the ability to use patient-paired eEC and eSF in experiments. These methods have been previously published, but here the protocol has been updated for maximum efficiency.
[摘要] 来自子宫内膜活检的子宫内膜上皮细胞(eEC)和基质成纤维细胞(eSF)的纯化和培养允许下游细胞特异性体外研究。 该协议的效用是细胞被净化而不受不想要的细胞类型的污染,以及在实验中使用患者配对的eEC和eSF的能力。 这些方法以前已经发布,但在这里协议已经更新为最大的效率。
|
|
Ex vivo Human Natural Killer (NK) Cell Stimulation and Intracellular IFNγ and CD107a Cytokine Staining
|
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。
|
|