An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization
An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization
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DOI：
10.3791/50332
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发表时间：
2013-06-01
影响因子：
1.2
通讯作者：
Gleissner, Christian A.
中科院分区：
文献类型：
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作者：
Erbel, Christian;Rupp, Gregor;Gleissner, Christian A.
单核细胞衍生的巨噬细胞代表先天免疫系统的重要细胞类型。研究巨噬细胞生物学的小鼠模型存在小鼠和人类单核细胞来源的巨噬细胞之间的表型和功能差异。因此，我们在这里描述了一种体外模型来生成和研究原代人类巨噬细胞。简而言之，对从前臂静脉抽取的外周血进行密度梯度离心后，使用负磁珠分离从外周血单核细胞中分离出单核细胞。然后将这些单核细胞在特定条件下培养六天，以诱导不同类型的巨噬细胞分化或极化。该模型易于使用，并且避免了小鼠和人之间物种特异性差异引起的问题。此外，它比使用永生化细胞系更接近体内条件。总之，这里描述的模型适合研究巨噬细胞生物学、识别疾病机制和新的治疗靶点。尽管不能完全取代用动物或死后获得的人体组织进行的实验，但这里描述的模型可以识别和验证可能与各种人类疾病高度相关的疾病机制和治疗靶点。
Monocyte-derived macrophages represent an important cell type of the innate immune system. Mouse models studying macrophage biology suffer from the phenotypic and functional differences between murine and human monocyte-derived macrophages. Therefore, we here describe an in vitro model to generate and study primary human macrophages. Briefly, after density gradient centrifugation of peripheral blood drawn from a forearm vein, monocytes are isolated from peripheral blood mononuclear cells using negative magnetic bead isolation. These monocytes are then cultured for six days under specific conditions to induce different types of macrophage differentiation or polarization. The model is easy to use and circumvents the problems caused by species-specific differences between mouse and man. Furthermore, it is closer to the in vivo conditions than the use of immortalized cell lines. In conclusion, the model described here is suitable to study macrophage biology, identify disease mechanisms and novel therapeutic targets. Even though not fully replacing experiments with animals or human tissues obtained post mortem, the model described here allows identification and validation of disease mechanisms and therapeutic targets that may be highly relevant to various human diseases.