不同方法诱导THP-1细胞分化效果比较

发布时间：2018-04-12 06:36

本文选题：THP- + 佛波酯　；参考：《中国比较医学杂志》2017年09期

【摘要】：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。
[Abstract]:Objective to optimize different methods to stimulate THP-1 cells to differentiate into M _ 1M _ 2 macrophages and DC cells, and to lay a foundation for the study of M _ 1M _ 2 and DC cell models in vitro.Methods at first, PMA and GM-CSF/M-CSF were used to stimulate the differentiation of THP-1 cells, and then different cytokines were added to induce the differentiation into M1M 2 and DC cells respectively. The morphological changes of the cells were observed and the expression of cell surface molecules was detected by flow cytometry.Results the expression of CD80 and CD86 on the surface of THP-1-M1 cells were significantly increased by the two methods. The expression of CD163 in THP-1-M2 cells and the expression of CD14 in THP-1-M2 cells decreased significantly.Overexpression of CD80, CD86 and CD11c.PMA stimulated M1M1M 2 and DC cells both adherent growth and GM-CSF / M-CSF stimulation, only partial adherent growth of DC cells, M 1 and M 2 cells, still showed suspension growth.Conclusion both methods can successfully induce the differentiation of THP-1 cells into different cell subtypes, but there are some differences in morphology of the induced cells, so we can choose the stimulation method according to the experimental demand.
【作者单位】：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;
【基金】：国家自然科学基金青年科学基金项目(编号:81301437) 科技部重大专项(编号:2014ZX10001001-001-004,2014ZX10001001-002-006)
【分类号】：R392

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