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

作　　者：彭卓颖 李想 丛喆 薛婧 

机构地区：[1]北京协和医学院比较医学中心,中国医学科学院医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,新发再发传染病动物模型研究北京市重点实验室,北京100021

出　　处：《中国比较医学杂志》2017年第10期10-15,23,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金-青年科学基金项目(81301437);科技部重大专项(2014ZX10001001-001-004,2014ZX10001001-002-006)

摘　　要：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.

关 键 词：THP-1 巨噬细胞 佛波酯 流式细胞术 

分 类 号：R-33[医药卫生]