流式细胞术分析PMA诱导THP-1分化为巨噬细胞的表型特征

Phenotypic characterization of PMA-induced THP-1 macrophages analyzed by flow cytometry

目的用流式细胞术明确单独使用PMA诱导THP-1细胞分化而来的巨噬细胞的分化情况。方法首先用PMA刺激THP-1细胞分化为巨噬细胞,再分别使用LPS、IFN-γ和IL-6将细胞诱导为M1型巨噬细胞,用IL-4、IL-13和IL-6将细胞诱导为M2型巨噬细胞。显微镜下观察三种细胞的形态学改变,并用流式细胞术检测各细胞主要CD分子表达的差异。结果当THP-1分化为巨噬细胞后变为贴壁生长,细胞形态呈较规则的圆形或椭圆形;M1和M2细胞形态不规则,细胞突起较明显,细胞颗粒度较大。巨噬细胞表面与抗原提呈功能相关的CD分子的表达均较低,大部分呈阴性;而M1和M2细胞表面这些CD分子的表达均较高。结论单独使用PMA刺激THP-1细胞所分化而来的巨噬细胞抗原提呈能力较弱,基本处于静息状态。

Objective To identify the characteristics of the subtype of PMA-induced THP-1 macrophages by flow cytometry analysis. Methods THP-1 monocytic cells differentiate into macrophages promoted by PMA,then induced into M1 and M2 by adding different cytokines,such as LPS,IL-6 and IFN-γ for THP-1-M1,IL-4,IL-13 and IL-6 for THP-1-M2. Morphology of cells were observed under a microscope and the expression of CD14,CD68,CD16,CD80,CD86,CD163,CD206,CD209,CD83,CD1 a,CD11c,HLA-DR were detected by flow cytometry. Results The macrophages stimulated by PMA became adherent; THP-1-M1 and THP-1-M2 lost their spherical morphology,appeared more irregular with many obvious projections. The expression of CD83,CD1 a,CD11c,HLA-DR which had the function of antigen presenting on the surface of THP-1-Mφ were very low,and most of them were negative,but those of THP-1-M1 and THP-1-M2 were very high. Conclusions The macrophages differentiated from THP-1 stimulated by PMA are weak in antigen presenting function.