Ju Lili, Zeng Hui, Chen Yun, Wu Yanhong, Wang Beibei, Xu Qunyuan
Department of Neurobiology, Beijing Institute for Brain Disorders, Beijing Center of Neural Regeneration and Repair, Key Laboratory for Neurodegenerative Diseases of the Ministry of Education, Capital Medical University, Beijing, China.
Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China.
J Neurosci Res. 2015 Sep;93(9):1345-52. doi: 10.1002/jnr.23563. Epub 2015 Jun 6.
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-β1 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.
小胶质细胞是中枢神经系统中多功能的免疫效应细胞，对各种刺激敏感。用于分离小胶质细胞的不同方法可能会影响其某些特性，例如其极化状态。细胞分选方法对小胶质细胞极化状态的影响尚未得到研究。混合胶质细胞培养系统（MGCS）和磁性激活细胞分选（MACS）是两种常用于纯化小胶质细胞的方法。本研究比较了通过MGCS分离的小胶质细胞和通过MACS分离的小胶质细胞之间的免疫状态。我们发现，通过MGCS分离的小胶质细胞比通过MACS分离的小胶质细胞表现出更强的免疫激活状态。它们具有更高的吞噬能力以及与经典激活（M1）和替代激活（M2）相关的高水平标志物。此外，在混合胶质细胞的培养基中检测到高水平的M1型和M2型趋化因子（C-C基序）配体2和转化生长因子-β1。我们的结果表明，通过MGCS分离的小胶质细胞处于免疫激活状态，而通过MACS分离的小胶质细胞似乎更接近其原始的体内状态。因此，在神经病理学研究中，应仔细考虑取决于纯化方法的小胶质细胞免疫状态。
