In vitro models of differing macrophage functions are useful since human monocyte-derived macrophages are short-lived, finite and vary from donor to donor. Published protocols using the promonocytic cell line THP-1 have tended to result in cells that closely resemble classically-activated macrophages, differentiated in IFNγ and LPS. However, no protocol, to date, has fully recapitulated polarization of THP-1 to the M(IL-4) or M(IL-10) macrophage phenotypes seen when human monocyte-derived macrophages are exposed to each cytokine. Here we present protocols that can be used to prepare M(IL-4) polarized THP-1 that transcribe CCL17, CCL26, CD200R and MRC1 and M(IL-10) cells which transcribe CD163, C1QA and SEPP1. We show that the inhibitory Fcγ Receptor IIb is preferentially expressed on the surface of M(IL-4) cells, altering the balance of activating to inhibitory Fcγ Receptors. Adoption of standardized experimental conditions for macrophage polarization will make it easier to compare downstream effector functions of different macrophage polarization states, where the impact of PMA exposure is minimized and rest periods and cytokine exposure have been optimized.

中文翻译：

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将 THP-1 细胞分化为具有不同 M(IFNγ+LPS)、M(IL-4) 和 M(IL-10) 表型的巨噬细胞的标准化方案。

不同巨噬细胞功能的体外模型很有用，因为人类单核细胞衍生的巨噬细胞寿命短、有限且因供体而异。使用促单核细胞系 THP-1 的已发布方案往往会产生与经典激活的巨噬细胞非常相似的细胞，这些细胞在 IFNγ 和 LPS 中分化。然而，迄今为止，没有任何协议完全概括了 THP-1 与 M(IL-4) 或 M(IL-10) 巨噬细胞表型的极化，当人类单核细胞衍生的巨噬细胞暴露于每种细胞因子时。在这里，我们提出可用于制备转录 CCL17、CCL26、CD200R 和 MRC1 的 M（IL-4）极化 THP-1 和转录 CD163、C1QA 和 SEPP1 的 M（IL-10）细胞的协议。我们表明抑制性 Fcγ 受体 IIb 优先在 M(IL-4) 细胞表面表达，改变激活和抑制 Fcγ 受体的平衡。采用标准化的巨噬细胞极化实验条件将使比较不同巨噬细胞极化状态的下游效应器功能变得更加容易，其中 PMA 暴露的影响最小化，休息时间和细胞因子暴露得到优化。