A phased strategy to differentiate human CD14+monocytes into classically and alternatively activated macrophages and dendritic cells.
A phased strategy to differentiate human CD14+monocytes into classically and alternatively activated macrophages and dendritic cells.
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DOI：
10.2144/000114435
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发表时间：
2016
期刊：
影响因子：
2.7
通讯作者：
Pienta KJ
中科院分区：
文献类型：
--
作者：
Zarif JC;Hernandez JR;Verdone JE;Campbell SP;Drake CG;Pienta KJ
目前有几种体外策略可以将从外周血单核细胞分离的人CD14+单核细胞分化为M1或M2巨噬细胞类型。然后，使用细胞表面标记的流式细胞术分析来验证每种细胞类型。人CD14+单核细胞具有向M1和M2巨噬细胞分化的潜能，两者都表现出不同程度的细胞表面抗原重叠。使用当前巨噬细胞极化方案的多个表面标记，我们的数据揭示了当前方法的几个局限性，例如具有细胞表面标记重叠和功能相似性的高度模糊的细胞类型。通过利用白介素6和两个阶段的细胞因子暴露，我们开发了一种方案，可以将人单核细胞分化为M1、M2或树突状细胞，相对于通常使用的方法产生的巨噬细胞和树突状细胞更有效和更逼真。这是通过改变细胞因子组成、剂量、孵育时间和改进验证方法来实现的。本研究为体外复制人单核细胞来源的树突状细胞和巨噬细胞模型提供了可靠的方法，有助于更好地定义和理解先天免疫和获得性免疫以及几种病理状态。
There are currently several in vitro strategies to differentiate human CD14+ monocytes isolated from peripheral blood mononuclear cells into M1 or M2 macrophage cell types. Thereafter, each cell type is then verified using flow cytometric analysis of cell surface markers. Human CD14+ monocytes have the potential to differentiate into M1 and M2 macrophages, both of which demonstrate varying degrees of cell surface antigen overlap. Using multiple surface markers with current macrophage polarization protocols, our data reveal several limitations with current methods, such as highly ambiguous cell types that possess cell surface marker overlap and functional similarities. By utilizing interleukin-6 and two phases of cytokine exposure, we have developed a protocol to differentiate human monocytes into M1, M2, or dendritic cells with a greater efficiently and fidelity relative to macrophages and dendritic cells that are produced from commonly used methods. This is achieved via alterations in cytokine composition, dosing, incubation times, and improvements in verification methodology. This report provides a reliable method to reproduce human in vitro monocyte derived dendritic cells and macrophage models that will aid in defining and understanding innate and adaptive immunity better as well as several pathologic states.
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影响因子：
4.4
作者：
Sallusto, Federica;Lanzavecchia, Antonio
通讯作者：
Lanzavecchia, Antonio
DOI：
10.1084/jem.19.3.283
发表时间：
1914-03-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Evans HM;Bowman FB;Winternitz MC
通讯作者：
Winternitz MC
DOI：
10.1073/pnas.93.6.2588
发表时间：
1996-03-19
影响因子：
11.1
作者：
Zhou, LJ;Tedder, TF
通讯作者：
Tedder, TF
DOI：
10.1084/jem.52.2.279
发表时间：
1930-07-31
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Forkner CE
通讯作者：
Forkner CE
基于甘露糖功能化苝双酰亚胺的蛋白质荧光开启传感及其荧光成像
影响因子：
12.6
作者：
Wang, Ke-Rang;An, Hong-Wei;Li, Xiao-Liu
通讯作者：
Li, Xiao-Liu