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In Vitro Generation of Monocyte-Derived Macrophages under Serum-Free Conditions Improves Their Tumor Promoting Functions

机译:在体外代单核细胞源性下无血清条件提高巨噬细胞的肿瘤推动作用

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摘要

The tumor promoting role of M2 macrophages has been described in in vivo models and the presence of macrophages in certain tumor types has been linked to a poor clinical outcome. In light of burgeoning activities to clinically develop new therapies targeting tumor-associated macrophages (TAMs), reliable in vitro models faithfully mimicking the tumor promoting functions of TAMs are required. Generation and activation of human monocyte-derived macrophages (MDM) in vitro, described as M1 or M2 macrophages attributed with tumoricidal or tumor-promoting functions, respectively, has been widely reported using mainly serum containing culture methods. In this study, we compared the properties of macrophages originating from monocytes cultured either in media containing serum together with M-CSF for M2 and GM-CSF for M1 macrophages or in serum-free media supplemented with M-CSF or GM-CSF and cytokines such as IL-4, IL-10 to induce activated M2 or LPS together with IFN-γ to generate activated M1 phenotype. We observed differences in cell morphology as well as increased surface receptor expression levels in serum-containing culture whereas similar or higher cytokine production levels were detected under serum-free culture conditions. More importantly, MDM differentiated under serum-free conditions displayed enhanced tumoricidal activity for M1 and tumor promoting property for M2 macrophages in contrast to MDM differentiated in the presence of serum. Moreover, evaluation of MDM phagocytic activity in serum free condition resulted in greater phagocytic properties of M2 compared to M1. Our data therefore confirm the tumor promoting properties of M2 macrophages in vitro and encourage the targeting of TAMs for cancer therapy.
机译:在体内模型中已经描述了M2巨噬细胞的肿瘤促进作用,并且在某些肿瘤类型中巨噬细胞的存在与不良的临床结果有关。鉴于迅速发展的临床开发针对肿瘤相关巨噬细胞(TAM)的新疗法的活动,需要忠实地模拟TAM促进肿瘤功能的可靠体外模型。广泛报道了体外人类单核细胞衍生巨噬细胞(MDM)的生成和激活,分别被描述为具有杀肿瘤或促肿瘤功能的M1或M2巨噬细胞,主要使用含血清的培养方法。在这项研究中,我们比较了在含有血清以及M1的M-CSF培养基和M1巨噬细胞的GM-CSF或补充M-CSF或GM-CSF和细胞因子的无血清培养基中培养的单核细胞巨噬细胞的特性。例如IL-4,IL-10诱导活化的M2或LPS以及IFN-γ产生活化的M1表型。我们观察到细胞形态的差异以及含血清培养物中表面受体表达水平的提高,而在无血清培养条件下检测到相似或更高的细胞因子产生水平。更重要的是,与无血清条件下分化的MDM相比,在无血清条件下分化的MDM对M1表现出增强的杀肿瘤活性,对M2巨噬细胞具有促进肿瘤的作用。此外,在无血清条件下对MDM吞噬活性的评估导致M2的吞噬特性高于M1。因此,我们的数据证实了M2巨噬细胞在体外的促肿瘤特性,并鼓励以TAM为靶点进行癌症治疗。

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