A new method to isolate microglia from adult mice and culture them for an extended period of time
A new method to isolate microglia from adult mice and culture them for an extended period of time
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DOI：
10.1016/j.jneumeth.2010.01.017
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发表时间：
2010-03-30
影响因子：
3
通讯作者：
Draheim, Henning Joerg
中科院分区：
文献类型：
--
作者：
Moussaud, Simon;Draheim, Henning Joerg
作为大脑的主要免疫活性细胞，小胶质细胞与神经保护和神经变性密切相关。因此，它们对于许多中枢神经系统疾病的研究具有重要意义。目前，为了模拟大脑炎症，广泛使用从胚胎或新生啮齿动物制备的小胶质细胞系或原代小胶质细胞。然而，这些体外小胶质细胞模型不适合神经退行性疾病领域的研究，其中衰老是一个关键参数。迄今为止，只有少数关于衰老小胶质细胞的体外研究发表，其中大多数使用不能长时间培养的离体小胶质细胞。在本研究中，我们提供了一种新方法，可以从成年小鼠大脑中分离和培养几乎纯的小胶质细胞群。分离基于结合密度分离和随后的培养物选择过程的程序。经过这些步骤后，小胶质细胞形成非粘附的漂浮细胞层，可以轻松、重复地收获和重新铺板。该方法简单，并且成体小胶质细胞的产量和纯度相对较高。收集的原代成体小胶质细胞增殖并可以在培养物中保存较长时间。我们将原代成年小胶质细胞与新生小鼠的原代小胶质细胞以及 C8-B4 小胶质细胞系进行了比较。我们发现成年小胶质细胞与其他体外模型具有相似但不相同的免疫表型、功能和电生理特征。 (C) 2010 Elsevier B.V. 保留所有权利。
As the major immuno-competent cells of the brain, microglia are highly implicated in neuro-protection as well as in neurodegeneration. Therefore, they are of key interest for research on numerous CNS diseases.Currently, to model inflammation in the brain, microglial cell lines or primary microglia prepared from embryonic or neo-natal rodents are widely used. However, these in vitro microglial models are not suitable for research in the field of neuro-degenerative diseases where aging is a crucial parameter. Only a few in vitro studies on aged microglia have been published so far, most of which use ex vivo microglia which cannot be kept in culture for prolonged periods of time.In the present study, we provide a new approach which allows the isolation and culture of an almost pure population of microglia from adult mouse brains. The isolation is based on a procedure which combines density separation and a subsequent culture selection process. After these steps, microglia form a non-adherent floating cell layer that can be easily and repeatedly harvested and replated. This method is simple and allows for a comparatively high yield and purity of adult microglial cells.The collected primary adult microglia proliferate and can be kept in culture for extended periods of time. We compared the primary adult microglia to primary microglia from neo-natal mice as well as to the C8-B4 microglial cell line. We found that adult microglia have similar, but not identical, immuno-phenotypic, functional and electrophysiological characteristics to the other in vitro models. (C) 2010 Elsevier B.V. All rights reserved.