Oxygen tension modulates differentiation and primary macrophage functions in the human monocytic THP-1 cell line.
Oxygen tension modulates differentiation and primary macrophage functions in the human monocytic THP-1 cell line.
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DOI：
10.1371/journal.pone.0054926
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lein PJ
中科院分区：
文献类型：
--
作者：
Grodzki AC;Giulivi C;Lein PJ
人THP-1细胞系被广泛用作巨噬细胞分化和功能研究的体外模型系统。这些细胞的常规培养条件包括环境氧压（~ 20% v/v）和添加硫醇-巯基乙醇（2-ME）和血清的培养基。考虑到O2和2-ME的氧化还原活性，以及大量支持活性氧（ROS）在巨噬细胞分化和功能中的作用的实验证据，我们解决了在没有2-ME和血清的情况下，在更生理相关的氧张力（5% O2）下培养THP-1细胞是否会改变THP-1细胞生理的问题。在18% O2和5% O2中培养的比较表明，降低氧张力对未分化THP-1细胞的增殖没有影响。然而，将氧张力降低至5% O2可显著提高酚酯诱导的THP-1细胞向巨噬细胞样细胞分化的速率，以及未分化和pma分化的THP-1细胞的代谢活性。从培养基中去除2-ME和血清会降低未分化THP-1细胞的增殖，但会增加代谢活性和两种氧张力下的分化率。在分化的THP-1细胞中，将氧张力降低至5% O2可降低吞噬活性、β-己糖氨酸酶的组成性释放和lps诱导的NF-κB活化，但可增强lps刺激的细胞因子释放。综上所述，这些数据表明，氧张力影响THP-1细胞分化和原代巨噬细胞功能，并提示在没有外源性还原剂和血清的情况下，在严格调节的氧张力下培养这些细胞，可能为研究局部氧化还原环境在调节THP-1细胞生理中的作用提供一个生理学相关的基线。
The human THP-1 cell line is widely used as an in vitro model system for studying macrophage differentiation and function. Conventional culture conditions for these cells consist of ambient oxygen pressure (∼20% v/v) and medium supplemented with the thiol 2-mercaptoethanol (2-ME) and serum. In consideration of the redox activities of O2 and 2-ME, and the extensive experimental evidence supporting a role for reactive oxygen species (ROS) in the differentiation and function of macrophages, we addressed the question of whether culturing THP-1 cells under a more physiologically relevant oxygen tension (5% O2) in the absence of 2-ME and serum would alter THP-1 cell physiology. Comparisons of cultures maintained in 18% O2 versus 5% O2 indicated that reducing oxygen tension had no effect on the proliferation of undifferentiated THP-1 cells. However, decreasing the oxygen tension to 5% O2 significantly increased the rate of phorbol ester-induced differentiation of THP-1 cells into macrophage-like cells as well as the metabolic activity of both undifferentiated and PMA-differentiated THP-1 cells. Removal of both 2-ME and serum from the medium decreased the proliferation of undifferentiated THP-1 cells but increased metabolic activity and the rate of differentiation under either oxygen tension. In differentiated THP-1 cells, lowering the oxygen tension to 5% O2 decreased phagocytic activity, the constitutive release of β-hexosaminidase and LPS-induced NF-κB activation but enhanced LPS-stimulated release of cytokines. Collectively, these data demonstrate that oxygen tension influences THP-1 cell differentiation and primary macrophage functions, and suggest that culturing these cells under tightly regulated oxygen tension in the absence of exogenous reducing agent and serum is likely to provide a physiologically relevant baseline from which to study the role of the local redox environment in regulating THP-1 cell physiology.
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DOI：
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