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Microglia pp 127–147Cite as

Replenishment of Organotypic Hippocampal Slice Cultures with Neonatal or Adult Microglia

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2034))

Abstract

This protocol describes a method to deplete and repopulate organotypic hippocampal slice cultures with ramified microglia. We describe the slice culture preparation from newborn mice, standard culturing of neonatal microglia, and the acute isolation of microglia from adult mouse brain. Furthermore, we outline the technique for the replenishment of microglia-depleted slice cultures with different microglia populations and subsequent morphological analysis. We show that neonatal and adult microglia acquire specific ramified morphologies, which in case of adult microglia are indistinguishable from the in vivo situation. This procedure not only allows the functional investigation of microglia with different degrees of ramification but also enables the construction of chimeric slice cultures with respect to the microglia phenotype. Preparation of slice cultures can be completed in 3.5 h, preparation of mixed-glial cultures in 4 h, isolation of adult microglia can be accomplished in 3.5 h, and replenishment in 30 min.

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Author information

Authors and Affiliations

  1. Institute of Clinical Chemistry and Laboratory Medicine, University Medicine Greifswald, Greifswald, Germany

    Annette Masuch

  2. Department of Psychiatry and Psychotherapy, University Hospital Freiburg, University of Freiburg, Freiburg, Germany

    Knut Biber

Authors
  1. Annette Masuch
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  2. Knut Biber
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Editor information

Editors and Affiliations

  1. Department of Neurophysiology, Institute of Physiology, Eberhard Karls University of Tübingen, Tübingen, Germany

    Olga Garaschuk

  2. Faculty of Biology, Medicine and Health The University of Manchester, , Center for Basic and Translational Neuroscience, Faculty of Health and Medical Sciences University of Copenhagen Copenhagen, Denmark, Achucarro Center for Neuroscience IKERBASQUE Basque Foundation for Science Bilbao, Spain, Manchester, UK

    Alexei Verkhratsky

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Masuch, A., Biber, K. (2019). Replenishment of Organotypic Hippocampal Slice Cultures with Neonatal or Adult Microglia. In: Garaschuk, O., Verkhratsky, A. (eds) Microglia. Methods in Molecular Biology, vol 2034. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9658-2_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9658-2_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9657-5

  • Online ISBN: 978-1-4939-9658-2

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