不同方法诱导THP-1细胞分化效果比较

Comparison of the effects of different treatments on THP-1 cell differentiation

目的优化不同方法刺激THP-1细胞定向分化为M1、M2巨噬细胞及DC细胞,为M1、M2和DC三种体外细胞模型研究奠定基础。方法首先用PMA和GM-CSF/M-CSF两种方法刺激诱导THP-1细胞分化,再分别添加不同细胞因子诱导其分化为M1、M2和DC细胞,观察细胞形态的变化,并用流式细胞术检测细胞表面分子的表达情况。结果两种方法刺激细胞CD分子表达的整体趋势基本一致。THP-1-M1细胞表面CD80和CD86表达量显著增加;THP-1-M2细胞高表达CD163和CD209;THP-1-DC细胞CD14表达量显著降低,高表达CD80、CD86和CD11c。PMA刺激后,M1、M2和DC细胞均贴壁生长;GM-CSF/M-CSF刺激后,只有DC细胞部分贴壁生长,M1和M2细胞仍呈悬浮生长。结论两种方法均能成功地诱导THP-1细胞向不同细胞亚型分化,但是诱导出来的细胞在形态上存在一定差异,可根据实验需求选择刺激方法。

Objective To stimulate a human monocytic cell line THP-1 cells to differentiate into M1, M2 macro-phages and dendritic （DC） cells by optimization of different methods, and lay the foundation for the study of M1, M2 and DC cell models in vitro. Methods THP-1 cells were stimulated by PMA and GM-CSF/M-CSF, respectively. Then, they were induced to differentiate into M1, M2 macrophages and DC cells by adding different cytokines, such as LPS, IL-6 and IFN-γ for M1 macrophages, IL-4, IL-13 and IL-6 for M2 macrophages, and IL-4 for DCs. Subsequently, the morphology of cells was observed and the expression of cell surface （ CD） molecules was detected by flow cytometry. Results After stimulation with the two methods, the trends of CD molecules expression were basically the same. The expression of CD80 and CD86 on the THP-1-M1 cells were increased significantly, and CD163 and CD209 were highly expressed on the THP-1-M2 cells. For THP-1-DC cells, the expression of CD14 was significantly decreased, while the expression of CD80, CD86 and CD11c increased. M1, M2 macrophages and DC cells were adherent after stimulation with PMA. However, DC cells were partially adherent after GM-CSF/M-CSF treatment. M1 and M2 macrophages were also growing in suspension. Conclusions Both methods used in this study can successfully induce THP-1 cells to differentiate into different subtypes, but there are some differences in the morphology of the induced cells. Appropriate stimulation method can be selected ac-cording to the experimental requirements.