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An optimized protocol for the acute isolation of human microglia from autopsy brain samples

Authors

  • Marta Olah,

    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
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    • Marta Olah and Divya Raj have contributed equally to the mansucript.

  • Divya Raj,

    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
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    • Marta Olah and Divya Raj have contributed equally to the mansucript.

  • Nieske Brouwer,

    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
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  • Alexander H. De Haas,

    1. Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands
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  • Bart J.L. Eggen,

    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
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  • Wilfred F. A. Den Dunnen,

    1. Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands
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  • Knut P. H. Biber,

    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
    2. Section Molecular Psychiatry, Department of Psychiatry and Psychotherapy, University Medical Center Freiburg, Freiburg, Germany
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  • Hendrikus W. G. M. Boddeke

    Corresponding author
    1. Section Medical Physiology, Department of Neuroscience, UMCG–RuG, Groningen, The Netherlands
    • Department of Medical Physiology, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands
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Abstract

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. © Wiley Periodicals, Inc.

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