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Author:
Date:
2020-06-05
[Abstract] The study of human neutrophils in vitro is challenging due to their short half-life and propensity for activation. However, with careful handling and manipulation in the laboratory, they can be a powerful tool to investigate immune responses in health and disease. Here we describe a method for the isolation of human neutrophils from peripheral blood samples, followed by a high-throughput screen to assess the efficacy of a library of compounds in inducing neutrophil apoptosis, which may have therapeutic potential in neutrophil-driven diseases. This protocol is based on previously-published neutrophil isolation methods utilizing Dextran sedimentation of red blood cells followed by the separation of granulocytes with plasma/Percoll discontinuous gradient centrifugation. Yields of ~1 ...
[摘要] [摘要] 人类嗜中性粒细胞的研究 由于其半衰期短且易于活化，因此体外具有挑战性，然而，通过在实验室中的仔细处理和操纵，它们可以成为研究健康和疾病中免疫反应的有力工具。在此，我们介绍了一种分离方法外周血样本中的人类嗜中性粒细胞，然后进行高通量筛选，以评估化合物库诱导嗜中性粒细胞凋亡的功效，该化合物在嗜中性粒细胞驱动的疾病中可能具有治疗潜力。此规程基于先前发表的嗜中性粒细胞分离方法利用红细胞的葡聚糖沉降，接着用等离子体/粒细胞的分离的Percoll 〜1×10个的不连续梯度centrifugation.Yields 6 每嗜中性粒细胞毫升的血液，和>的纯度95个％嗜中性粒细胞是典型的。嗜中性粒细胞与经处理的激酶抑制剂文库，然后通过流式细胞仪评估中性粒细胞凋亡的速率。低通量用于高通量筛选人类原代免疫细胞，以鉴定具有修饰嗜中性粒细胞功能的化合物，并可根据需要进行修饰以评估其他表型。
[背景] 中性粒细胞是重要的先天免疫细胞与键的角色防御抵抗infection.They是短暂的细胞中，在感染和inflammat部位延长6-8小时循环典型的半衰期离子（萨默斯等人。 。，2010 ; Hidalgo。Et Al ...
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