Department of Immunology, Max Rady College of Medicine, Rady Faculty of Health Science, University of Manitoba, Winnipeg, MB, Canada.
Children's Hospital Research Institute of Manitoba, University of Manitoba, Winnipeg, MB, Canada.
Methods Mol Biol. 2020;2184:131-144. doi: 10.1007/978-1-0716-0802-9_10.
Macrophages are professional innate immune cells that are broadly disseminated throughout the body, shape various innate and adaptive immune responses, and play crucial roles in inflammation, homeostasis, wound healing, and tissue remodelling. According to their surrounding microenvironments, macrophages can differentiate themselves in different phenotypes. Over the last two decades, gene expression profiling has been used to decipher new transcripts associated with macrophage phenotypes. This chapter outlines protocols used to isolate and culture murine macrophages and how they can be "polarized" to obtain a specific phenotype. Furthermore, we describe a protocol for gene expression profiling using a quantitative real-time polymerase chain reaction (qPCR), a high-standard technology in the field of gene expression.
巨噬细胞是广泛分布于全身的专业先天免疫细胞，它们可以调节各种先天和适应性免疫反应，并在炎症、稳态、伤口愈合和组织重塑中发挥关键作用。根据周围的微环境，巨噬细胞可以分化成不同的表型。在过去的二十年中，基因表达谱分析已被用于破译与巨噬细胞表型相关的新转录本。本章概述了用于分离和培养鼠巨噬细胞的方案，以及如何“极化”它们以获得特定的表型。此外，我们还描述了一种使用定量实时聚合酶链反应（qPCR）进行基因表达谱分析的方案，qPCR 是基因表达领域的一项高标准技术。
