Vong Linda, Sherman Philip M, Glogauer Michael
Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
Methods Mol Biol. 2013;1031:41-50. doi: 10.1007/978-1-62703-481-4_5.
Neutrophils are one of the first cells to respond to an inflammatory stimulus, and are equipped with an assortment of antimicrobial and proteolytic enzymes to disarm and degrade bacterial pathogens. A novel mechanism of bacterial trapping, termed neutrophil extracellular traps (NETs), was recently described whereby neutrophils were shown to cast out web-like structures of chromatin, capturing and immobilizing invading pathogens. Herein we describe protocols to isolate murine bone marrow-derived neutrophils, and spectrophotometrically quantify, immunolabel, and visualize NET structures in vitro.
中性粒细胞是最早对炎症刺激作出反应的细胞之一,它们配备了各种抗菌和蛋白水解酶,以解除细菌病原体的武装并将其降解。最近描述了一种新的细菌捕获机制,称为中性粒细胞胞外陷阱(NETs),据此显示中性粒细胞会抛出染色质的网状结构,捕获并固定入侵的病原体。在此,我们描述了分离小鼠骨髓来源中性粒细胞的方法,以及通过分光光度法定量、免疫标记和体外观察NET结构的方法。