An optimized protocol for the acute isolation of human microglia from autopsy brain samples
An optimized protocol for the acute isolation of human microglia from autopsy brain samples
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DOI：
10.1002/glia.21251
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发表时间：
2012-01-01
期刊：
影响因子：
6.2
通讯作者：
Boddeke, Hendrikus W. G. M.
中科院分区：
文献类型：
--
作者：
Olah, Marta;Raj, Divya;Boddeke, Hendrikus W. G. M.
人们越来越认识到小胶质细胞在维持大脑和脊髓的组织稳态中起着至关重要的作用。因此，与中枢神经系统的各种生理和病理过程相关的小胶质细胞表型的科学兴趣日益增长，这并不奇怪。直到最近，由于缺乏一种分离方案（没有延长的培养期）来提供高纯度和高产量的小胶质细胞群体，对这些表型的研究受到阻碍。因此，我们的目标是建立一种快速有效的方法从人死后脑样本中分离小胶质细胞。我们测试了现有方案的多个要素（例如，密度分离、免疫磁珠分离），并将它们结合起来，以最大限度地减少制备时间，最大限度地提高产量和纯度。本文中介绍的程序能够从尸检（和活检）样本中急性分离人类小胶质细胞，其纯度和产量适合下游应用，例如蛋白质和基因表达分析以及功能分析。此外，本方案适用于从尸检样本中分离小胶质细胞，而不考虑大脑或特定大脑区域的神经状态，并且（稍加修改）甚至可用于从人类胶质瘤组织中分离小胶质细胞。(c)威利期刊公司
Microglia are increasingly recognized to be crucially involved in the maintenance of tissue homeostasis of the brain and spinal cord. Not surprisingly is therefore the growing scientific interest in the microglia phenotypes associated with various physiological and pathological processes of the central nervous system. Until recently the investigation of these phenotypes was hindered by the lack of an isolation protocol that (without an extended culturing period) would offer a microglia population of high purity and yield. Thus, our objective was to establish a rapid and efficient method for the isolation of human microglia from postmortem brain samples. We tested multiple elements of already existing protocols (e.g., density separation, immunomagnetic bead separation) and combined them to minimize preparation time and maximize yield and purity. The procedure presented in this article enables acute isolation of human microglia from autopsy (and biopsy) samples with a purity and yield that is suitable for downstream applications, such as protein and gene expression analysis and functional assays. Moreover, the present protocol is appropriate for the isolation of microglia from autopsy samples irrespective of the neurological state of the brain or specific brain regions and (with minor modification) could be even used for the isolation of microglia from human glioma tissue. (c) Wiley Periodicals, Inc.