Bajpai Geetika, Lavine Kory J
Department of Medicine, Washington University School of Medicine.
Department of Medicine, Washington University School of Medicine; Department of Immunology and Pathology, Washington University School of Medicine; Department of Developmental Biology, Washington University School of Medicine;
J Vis Exp. 2019 Dec 17(154). doi: 10.3791/60015.
Macrophages represent the most heterogeneous and abundant immune cell populations in the heart and are central in driving inflammation and reparative responses after cardiac injury. How various subsets of macrophages orchestrate the immune responses after cardiac injury is an active area of research. Presented here is a simple protocol that our lab performs routinely, for the extraction of macrophages from mouse and human myocardium specimens obtained from healthy and diseased individuals. Briefly, this protocol involves enzymatic digestion of cardiac tissue to generate a single cell suspension, followed by antibody staining, and flow cytometry. This technique is suitable for functional assays performed on sorted cells as well as bulk and single cell RNA sequencing. A major advantage of this protocol is its simplicity, minimal day to day variation and wide applicability allowing investigation of macrophage heterogeneity across various mouse models and human disease entities.
巨噬细胞是心脏中最具异质性且数量丰富的免疫细胞群体，在心脏损伤后驱动炎症和修复反应中起核心作用。心脏损伤后，各种巨噬细胞亚群如何协调免疫反应是一个活跃的研究领域。本文介绍了我们实验室常规执行的一个简单方案，用于从健康和患病个体获取的小鼠和人心肌标本中提取巨噬细胞。简而言之，该方案包括对心脏组织进行酶消化以生成单细胞悬液，随后进行抗体染色和流式细胞术。这项技术适用于对分选细胞进行的功能测定以及批量和单细胞RNA测序。该方案的一个主要优点是其简单性、日常变化最小以及适用性广泛，可用于研究各种小鼠模型和人类疾病实体中的巨噬细胞异质性。
