The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus
The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus
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DOI：
10.1016/j.jim.2016.01.012
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发表时间：
2016-03-01
影响因子：
2.2
通讯作者：
Donnelly, Sheila
中科院分区：
文献类型：
--
作者：
Lund, Maria E.;To, Joyce;Donnelly, Sheila
人单核细胞系THP-1是原代人单核细胞/巨噬细胞最广泛使用的模型。这是因为，在使用佛波酯112-肉豆蔻酸酯13-乙酸酯（PMA）分化后，THP-1细胞获得巨噬细胞样表型，其在许多方面模仿原代人巨噬细胞。尽管THP-1细胞广泛用于阐明巨噬细胞对炎性刺激的反应的研究以及潜在治疗剂的开发和筛选，但目前还没有使用PMA将THP-1单核细胞可靠分化为巨噬细胞表型的标准化方案。因此，使用THP-1细胞的报道已经证明所得THP-1巨噬细胞群体之间存在显著的表型和功能差异，这主要归因于所使用的不同PMA分化方法。因此，为了保证研究之间的一致性和再现性，并确保THP-1细胞作为原代人巨噬细胞的适当模型的相关性，开发THP-1巨噬细胞分化的标准化方案至关重要。因此，我们比较了使用一系列公开的PMA分化方案产生的THP-1巨噬细胞的功能和表型，特别是响应于促炎刺激物脂多糖（LPS）。我们的研究结果表明，所得到的THP-1巨噬细胞群体的功能，确定由肿瘤坏死因子（TNF）分泌响应LPS刺激，变化显着，并依赖于PMA的浓度用于刺激单核细胞的分化，和PMA暴露后的休息时间。这些数据表明，单核细胞THP-1细胞暴露于25 nM PMA超过48小时，随后在不存在PMA的情况下培养24小时的恢复期，是THP-1细胞分化的最佳方案。皇冠版权所有（C）2016由Elsevier B. V.出版。保留所有权利。
The human monocytic cell line, THP-1, is the most widely used model for primary human monocytes/macrophages. This is because, following differentiation using phorbo112-myristate 13-acetate (PMA), THP-1 cells acquire a macrophage-like phenotype, which mimics, in many respects, primary human macrophages. Despite the widespread use of THP-1 cells in studies elucidating macrophage responses to inflammatory stimuli, as well as the development and screening of potential therapeutics, there is currently no standardised protocol for the reliable differentiation of THP-1 monocytes to a macrophage phenotype using PMA. Consequently, reports using THP-1 cells have demonstrated significant phenotypic and functional differences between resultant THP-1 macrophage populations, which are largely attributable to the varying PMA differentiation methods used. Thus, to guarantee consistency and reproducibility between studies, and to ensure the relevance of THP-1 cells as an appropriate model for primary human macrophages, it is crucial to develop a standardised protocol for the differentiation of THP-1 macrophages. Accordingly, we compared the function and phenotype of THP-1 macrophages generated using the range of published PMA differentiation protocols, specifically in response to the pro inflammatory stimulus, lipopolysaccharide (LPS). Our results demonstrated that the function of the resultant THP-1 macrophage populations, as determined by tumour necrosis factor (TNF) secretion in response to LPS stimulation, varied significantly, and was dependent upon the concentration of PMA used to stimulate the differentiation of monocytes, and the period of rest following PMA exposure. These data indicate that exposure of monocytic THP-1 cells to 25 nM PMA over 48 h, followed by a recovery period of 24 h in culture in the absence of PMA, was the optimal protocol for the differentiation of THP-1 cells. Crown Copyright (C) 2016 Published by Elsevier B.V. All rights reserved.