Production and Characterization of Human Macrophages from Pluripotent Stem Cells.
Production and Characterization of Human Macrophages from Pluripotent Stem Cells.
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DOI：
10.3791/61038
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发表时间：
2020-04
期刊：
影响因子：
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通讯作者：
Martha Lopez-Yrigoyen;Alisha May;T. Ventura;H. Taylor;A. Fidanza;Luca Cassetta;J. Pollard;L. Forrester
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文献类型：
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作者：
Martha Lopez-Yrigoyen;Alisha May;T. Ventura;H. Taylor;A. Fidanza;Luca Cassetta;J. Pollard;L. Forrester
巨噬细胞存在于大多数脊椎动物组织中，包括具有不同功能的广泛分散和异质的细胞群。它们是健康和疾病的关键参与者，在免疫防御过程中充当吞噬细胞，并介导营养，维护和修复功能。虽然已经有可能研究参与人类巨噬细胞功能的一些分子过程，但已证明难以将基因工程技术应用于初级人类巨噬细胞。这大大阻碍了我们询问巨噬细胞生物学中涉及的复杂遗传途径和生成特定疾病状态模型的能力。因此，一种现成的人类巨噬细胞来源，可以适用于大量的遗传操作技术，将在这一领域提供一个有价值的工具。我们提出了一种优化的方案，允许在体外从人诱导多能干细胞（iPSC）产生巨噬细胞。这些iPSC衍生的巨噬细胞（iPSC-DM）表达人巨噬细胞表面标志物，包括CD 45，25 F9，CD 163和CD 169，我们的活细胞成像功能测定表明它们表现出强大的吞噬活性。培养的iPSC-DM可通过添加LPS和IFNg、IL 4或IL 10而被活化至显示改变的基因表达和吞噬活性的不同巨噬细胞状态。因此，该系统提供了一个平台来产生携带模拟特定人类疾病的遗传改变的人巨噬细胞，以及用于治疗这些疾病的药物筛选或细胞疗法的细胞来源。
Macrophages are present in most vertebrate tissues and comprise widely dispersed and heterogeneous cell populations with different functions. They are key players in health and disease, acting as phagocytes during immune defense and mediating trophic, maintenance, and repair functions. Although it has been possible to study some of the molecular processes involved in human macrophage function, it has proved difficult to apply genetic engineering techniques to primary human macrophages. This has significantly hampered our ability to interrogate the complex genetic pathways involved in macrophage biology and to generate models for specific disease states. An off-the-shelf source of human macrophages that is amenable to the vast arsenal of genetic manipulation techniques would, therefore, provide a valuable tool in this field. We present an optimized protocol that allows for the generation of macrophages from human induced pluripotent stem cells (iPSCs) in vitro. These iPSC-derived macrophages (iPSC-DMs) express human macrophage cell surface markers, including CD45, 25F9, CD163, and CD169, and our live-cell imaging functional assay demonstrates that they exhibit robust phagocytic activity. Cultured iPSC-DMs can be activated to different macrophage states that display altered gene expression and phagocytic activity by the addition of LPS and IFNg, IL4, or IL10. Thus, this system provides a platform to generate human macrophages carrying genetic alterations that model specific human disease and a source of cells for drug screening or cell therapy to treat these diseases.