Isolation of Mouse Neutrophils
Isolation of Mouse Neutrophils
复制标题
DOI：
10.1002/cpz1.879
复制
发表时间：
1997-06
期刊：
影响因子：
--
通讯作者：
M. Swamydas;Yi Luo;M. Dorf;M. Lionakis
中科院分区：
文献类型：
--
作者：
M. Swamydas;Yi Luo;M. Dorf;M. Lionakis
中性粒细胞是抵抗细菌和真菌病原体的第一道防线。事实上，具有遗传性或获得性定性和定量中性粒细胞缺陷的患者处于发生细菌和真菌感染并遭受这些感染的不良后果的高风险中。因此，旨在确定在稳态条件下和感染期间调节中性粒细胞效应子功能的分子因子的研究对于设计增强中性粒细胞功能和改善感染个体的结果的策略至关重要。本文描述了可重复的基于密度梯度离心的以及阳性和阴性免疫磁性选择方案，可应用于任何实验室，从小鼠骨髓中收获大量高度富集和高活力的中性粒细胞。在另一个方案中，我们还提出了一种方法，该方法将温和的酶组织消化与阳性免疫磁性选择技术或荧光激活细胞分选（FACS）相结合，以直接从小鼠组织（如肾脏，肝脏或脾脏）中收获高纯度和高活力的中性粒细胞制剂。通过这些方案分离的小鼠嗜中性粒细胞可用于检查离体细胞功能的几个方面，包括病原体结合、吞噬作用和杀伤、嗜中性粒细胞趋化性、氧化爆发、脱粒和细胞因子产生，以及用于进行嗜中性粒细胞过继转移实验。© 2023 Wiley Periodicals LLC.本文由美国政府雇员贡献，他们的作品在美国属于公有领域。
Neutrophils represent the first line of defense against bacterial and fungal pathogens. Indeed, patients with inherited or acquired qualitative and quantitative neutrophil defects are at high risk for developing bacterial and fungal infections and suffering adverse outcomes from these infections. Therefore, research aiming at defining the molecular factors that modulate neutrophil effector function under homeostatic conditions and during infection is essential for devising strategies to augment neutrophil function and improve the outcomes of infected individuals. This article describes reproducible density‐gradient‐centrifugation‐based as well as positive and negative immunomagnetic selection protocols that can be applied in any laboratory to harvest large numbers of highly enriched and highly viable neutrophils from the bone marrow of mice. In another protocol, we also present a method that combines gentle enzymatic tissue digestion with a positive immunomagnetic selection technique or fluorescence‐activated cell sorting (FACS) to harvest highly pure and highly viable preparations of neutrophils directly from mouse tissues such as the kidney, the liver, or the spleen. Mouse neutrophils isolated by these protocols can be used to examine several aspects of cellular function ex vivo, including pathogen binding, phagocytosis, and killing, neutrophil chemotaxis, oxidative burst, degranulation, and cytokine production, and for performing neutrophil adoptive transfer experiments. © 2023 Wiley Periodicals LLC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.