从小鼠骨髓中生成并鉴定GM-CSF衍生的肺泡样巨噬细胞和树突状细胞
Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow.
作者信息
Dong Yifei, Arif Arif A, Poon Grace F T, Hardman Blair, Dosanjh Manisha, Johnson Pauline
机构信息
Microbiology and Immunology, University of British Columbia.
Microbiology and Immunology, University of British Columbia;
出版信息
J Vis Exp. 2016 Jun 25(112):54194. doi: 10.3791/54194.
Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. However, they are difficult to isolate in sufficient numbers to study them in detail, therefore, in vitro models have been developed. In vitro cultures of bone marrow-derived macrophages and dendritic cells are well-established and valuable methods for immunological studies. Here, a method for culturing and identifying both DCs and macrophages from a single culture of primary mouse bone marrow cells using the cytokine granulocyte macrophage colony-stimulating factor (GM-CSF) is described. This protocol is based on the established procedure first developed by Lutz et al. in 1999 for bone marrow-derived DCs. The culture is heterogeneous, and MHCII and fluoresceinated hyaluronan (FL-HA) are used to distinguish macrophages from immature and mature DCs. These GM-CSF derived macrophages provide a convenient source of in vitro derived macrophages that closely resemble alveolar macrophages in both phenotype and function.
巨噬细胞和树突状细胞（DCs）是存在于组织和淋巴器官中的固有免疫细胞，在抵御病原体方面发挥关键作用。然而，难以分离出足够数量的这些细胞来进行详细研究，因此，已开发出体外模型。骨髓来源的巨噬细胞和树突状细胞的体外培养是成熟且有价值的免疫学研究方法。在此，描述了一种使用细胞因子粒细胞巨噬细胞集落刺激因子（GM-CSF）从原代小鼠骨髓细胞的单一培养物中培养和鉴定DCs及巨噬细胞的方法。该方案基于Lutz等人于1999年首次开发的用于骨髓来源DCs的既定程序。培养物是异质性的，主要组织相容性复合体II类分子（MHCII）和荧光素化透明质酸（FL-HA）用于区分巨噬细胞与未成熟和成熟的DCs。这些GM-CSF衍生的巨噬细胞提供了一种方便的体外衍生巨噬细胞来源，其在表型和功能上都与肺泡巨噬细胞非常相似。
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