Rapid isolation and culture of primary microglia from adult mouse spinal cord
Rapid isolation and culture of primary microglia from adult mouse spinal cord
复制标题
DOI：
10.1016/j.jneumeth.2009.07.002
复制
发表时间：
2009-10-15
影响因子：
3
通讯作者：
Malcangio, Marzia
中科院分区：
文献类型：
--
作者：
Yip, Ping K.;Kaan, Timothy K. Y.;Malcangio, Marzia
小胶质细胞在动态平衡中很重要，被广泛认为在神经病理性疼痛和多发性硬化症的发病机制中发挥作用。研究成人脊髓中的小胶质细胞对于进一步了解这些细胞在疾病病理中的作用是必不可少的。初级小胶质细胞通常是从胚胎或围产期啮齿动物的脑组织中提取的，这个过程可能需要一周多的时间才能完成。这项研究中的方案提供了从成人脊髓中快速分离小胶质细胞的方法，允许在几个小时内立即用于体外和体外实验。纯度可达99%，几乎没有神经细胞或星形胶质细胞的污染。这些成年小胶质细胞中有70%到85%处于相对非激活状态。在功能上，与未刺激的小胶质细胞相比，这些小胶质细胞对脂多糖孵育的反应与磷酸化p38MAPK和OX42免疫染色的增加以及ATP的释放有关。该技术提供了一种快速有效地从成年小鼠脊髓中提取高纯度、静止和功能活跃的小胶质细胞的方法，从而可以更深入地研究成年小胶质细胞在生理和病理生理状态下的情况。(C)2009爱思唯尔B.V.保留所有权利。
Microglia are important in homeostasis and widely considered to have roles in the pathogenesis of conditions such as neuropathic pain and multiple sclerosis. The need to study microglia from the adult spinal cord is essential to further understand the role of these cells in disease pathology. Primary microglia are often prepared from brain tissues obtained from embryonic or perinatal age rodents and the process can take over a week to complete. The protocol in this study provides rapid isolation of microglia from adult spinal cord, allowing immediate availability for experimentation of both ex vivo and in vitro within a few hours. A purity of 99% with little or no neuronal or astrocytic contamination can be achieved. Between 70% and 85% of these adult microglia were in a relatively non-activated state. Functionally, these microglia respond to lipopolysaccharide incubation with increases in both phospho-p38 MAPK and OX42 immunostaining, as well as release of ATP, as compared to un-stimulated microglia. This technique provides a protocol to achieve rapid and efficient extraction of high purity, quiescent and functionally active microglia from adult mouse spinal cord, allowing greater study of adult spinal microglia in physiological and pathophysiological states. (C) 2009 Elsevier B.V. All rights reserved.