Optimized THP-1 differentiation is required for the detection of responses to weak stimuli
Optimized THP-1 differentiation is required for the detection of responses to weak stimuli
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DOI：
10.1007/s00011-007-6115-5
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发表时间：
2007-01-01
影响因子：
6.7
通讯作者：
Kim, K. S.
中科院分区：
文献类型：
--
作者：
Park, E. K.;Jung, H. S.;Kim, K. S.
目的：THP-1单核细胞向巨噬细胞的分化主要是在12-肉豆酸13-醋酸酯（PMA）浓度为10-400ng/ml的条件下进行的。然而，该浓度可能高到足以上调分化巨噬细胞中某些基因的表达，从而压倒其他刺激诱导的基因表达增加。本研究旨在优化分化单核细胞所需的PMA浓度，同时最小化基因上调。方法：用2.5 ~ 100 ng/ml PMA处理THP-1细胞，观察细胞黏附程度、巨噬细胞表面标记物、细胞稳定分化、无不良基因上调。用10 ng/ml LPS或125 nM β淀粉样蛋白（A β(1-42)）处理最低PMA浓度的THP-1细胞，发现5 ng/ml PMA处理THP-1足以诱导稳定分化，没有不良的基因上调。这些在5 ng/ml浓度下分化的巨噬细胞对10 ng/mI LPS或125 nM β淀粉样蛋白等次级弱刺激反应良好（A β(1-42)）。结论：提示5 ng/ml PMA可使THP-1细胞分化良好，分化后的巨噬细胞对继发性弱刺激反应良好，不会被PMA诱导的不良基因上调所淹没。
Objectives: The differentiation of THP-1 monocytes into macrophages is mainly conducted at a phorbol 12-myristate 13-acetate (PMA) concentration of 10-400ng/ml. However, this concentration might be high enough to upregulate the expressions of some genes in differentiated macrophages, which could overwhelm gene expression increases induced by other stimuli. The present study was performed to optimize the PMA concentration required to differentiate monocytes whilst minimizing gene upregulation.Methods: THP-1 cells were treated with 2.5-100 ng/ml PMA and analyzed for the extent of cell adherence, the surface marker of macrophages, and stable differentiation without undesirable gene upregulation. The stably differentiated THP-1 cells at the minimum PMA concentration were treated with 10 ng/ml LPS or 125 nM amyloid beta (A beta(1-42)),Results: The treatment of THP-1 with 5 ng/ml PMA was found to be sufficient to induce stable differentiation without undesirable gene upregulation. These macrophages differentiated at 5 ng/ml responded well to secondary weak stimuli like 10 ng/mI LPS or 125 nM of amyloid beta (A beta(1-42)).Conclusions: This finding suggests that THP-1 cells are well differentiated by 5 ng/ml PMA, and that the resulting differentiated macrophages respond well to secondary weak stimuli without being overwhelmed by undesirable gene upregulation induced by PMA.