Dual polarization of microglia isolated from mixed glial cell cultures
Dual polarization of microglia isolated from mixed glial cell cultures
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DOI：
10.1002/jnr.23563
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发表时间：
2015-09-01
影响因子：
4.2
通讯作者：
Xu, Qunyuan
中科院分区：
文献类型：
--
作者：
Ju, Lili;Zeng, Hui;Xu, Qunyuan
小胶质细胞是中枢神经系统的多功能免疫效应细胞，对各种刺激敏感。分离小胶质细胞的不同方法可能会影响它们的一些特性，例如它们的极化状态。细胞分选方法对小胶质细胞极化状态的影响从未被研究过。混合胶质细胞培养系统（MGCS）和磁激活细胞分选（MACS）是目前常用的两种纯化小胶质细胞的方法。本研究比较了MGCS法和MACS法分离的小胶质细胞的免疫状态。我们发现，MGCS分离的小胶质细胞表现出比MACS分离的小胶质细胞更强的免疫激活状态。它们表现出升高的吞噬能力和高水平的与经典活化（M1）和替代活化（M2）相关的标志物。此外，在混合胶质细胞的培养液中检测到高水平的M1型和M2型趋化因子（C-C基序）配体2和转化生长因子-1。我们的研究结果表明，MGCS分离的小胶质细胞处于免疫激活状态，而MACS分离的小胶质细胞似乎更接近其主要的体内状态。因此，在神经病理学研究中，应仔细考虑小胶质细胞的免疫状态，这取决于用于纯化它们的方案。(c)2015 Wiley Periodicals，Inc.
Microglia are versatile immune effector cells of the CNS and are sensitive to various stimuli. The different methods used to isolate microglia may affect some of their characteristics, such as their polarization state. The influence of cell sorting methods on the polarization state of microglia has never been studied. Mixed glial culture system (MGCS) and magnetic activated cell sorting (MACS) are two methods that are commonly used to purify microglia. This study compares the immunological states between microglia isolated by MGCS and microglia isolated by MACS. We show that microglia isolated by MGCS exhibit a stronger immune-activated state than microglia isolated by MACS. They present an elevated phagocytic ability and high levels of markers associated with classical activation (M1) and alternative activation (M2). In addition, high levels of M1-type and M2-type chemokine (C-C motif) ligand 2 and transforming growth factor-1 were detected in the culture medium of mixed glial cells. Our results show that microglia isolated by MGCS are in an immune-activated state, whereas microglia isolated by MACS appear to be closer to their primary in vivo state. Therefore, the immune status of microglia, depending on the protocol used to purify them, should be carefully considered in neuropathology research. (c) 2015 Wiley Periodicals, Inc.