小鼠骨髓来源中性粒细胞的中性粒细胞胞外陷阱（NETs）的定量与可视化
Quantification and Visualization of Neutrophil Extracellular Traps (NETs) from Murine Bone Marrow-Derived Neutrophils.
作者信息
Vong Linda, Sherman Philip M, Glogauer Michael
机构信息
Cell Biology Program, Research Institute, Hospital for Sick Children, University of Toronto, Toronto, ON, Canada.
Faculty of Dentistry, University of Toronto, Toronto, ON, Canada.
出版信息
Methods Mol Biol. 2019;1960:63-73. doi: 10.1007/978-1-4939-9167-9_5.
Neutrophils are some of the first leukocytes to respond to inflammatory stimuli. Once recruited, these cells are equipped with an assortment of proteolytic enzymes and antimicrobial factors that disarm and degrade pathogens. Neutrophils employ a highly novel mechanism to contain and trap bacteria in the local inflammatory microenvironment, termed neutrophil extracellular traps (NETs). During NET formation, neutrophils eject weblike structures of chromatin, which captures and immobilizes invading pathogens. In this chapter, we describe protocols to isolate bone marrow-derived neutrophils from mice. We further describe in vitro methods to spectrophotometrically quantify, immunolabel, and visualize NET structures.
中性粒细胞是最早对炎症刺激作出反应的白细胞之一。一旦被募集，这些细胞就配备了各种蛋白水解酶和抗菌因子，用于解除病原体的武装并将其降解。中性粒细胞采用一种非常新颖的机制，在局部炎症微环境中捕获和困住细菌，这一机制被称为中性粒细胞胞外诱捕网（NETs）。在NET形成过程中，中性粒细胞会排出染色质的网状结构，该结构能够捕获并固定入侵的病原体。在本章中，我们描述了从小鼠中分离骨髓源性中性粒细胞的方法。我们还进一步描述了通过分光光度法定量、免疫标记和可视化NET结构的体外方法。
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