Department of Neurology, University of Massachusetts Chan Medical School; Translation Science Program, Morningside Graduate School of Biomedical Sciences, University of Massachusetts Chan Medical School.
Department of Neurology, University of Massachusetts Chan Medical School;
J Vis Exp. 2022 Aug 18(186). doi: 10.3791/64323.
Microglia are the resident immune cells of myeloid origin that maintain homeostasis in the brain microenvironment and have become a key player in multiple neurological diseases. Studying human microglia in health and disease represents a challenge due to the extremely limited supply of human cells. Induced pluripotent stem cells (iPSCs) derived from human individuals can be used to circumvent this barrier. Here, it is demonstrated how to differentiate human iPSCs into microglia-like cells (iMGs) for in vitro experimentation. These iMGs exhibit the expected and physiological properties of microglia, including microglia-like morphology, expression of proper markers, and active phagocytosis. Additionally, documentation for isolating and labeling synaptosome substrates derived from human iPSC-derived lower motor neurons (iLMNs) is provided. A live-cell, longitudinal imaging assay is used to monitor engulfment of human synaptosomes labeled with a pH-sensitive dye, allowing for investigations of iMG's phagocytic capacity. The protocols described herein are broadly applicable to different fields that are investigating human microglia biology and the contribution of microglia to disease.
小胶质细胞是髓系来源的固有免疫细胞，它们在大脑微环境中维持着内环境稳态，并已成为多种神经疾病的关键参与者。由于人类细胞的极度缺乏，研究健康和疾病状态下的人类小胶质细胞是一个挑战。诱导多能干细胞（iPSCs）可来源于人类个体，可用于克服这一障碍。本文展示了如何将人类 iPSCs 分化为小胶质细胞样细胞（iMGs）进行体外实验。这些 iMGs 表现出小胶质细胞的预期和生理特性，包括小胶质细胞样形态、适当标志物的表达和活跃的吞噬作用。此外，还提供了分离和标记源自人类 iPSC 衍生的下运动神经元（iLMNs）的突触小体底物的文档。使用活细胞、纵向成像测定法监测用 pH 敏感染料标记的人突触小体的吞噬作用，从而可以研究 iMG 的吞噬能力。本文描述的方案广泛适用于研究人类小胶质细胞生物学和小胶质细胞对疾病贡献的不同领域。
