Microglia pp 127-147 | Cite as

Replenishment of Organotypic Hippocampal Slice Cultures with Neonatal or Adult Microglia

  • Annette Masuch
  • Knut Biber
Part of the Methods in Molecular Biology book series (MIMB, volume 2034)


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.

Key words

Organotypic hippocampal slice culture Microglia Mixed-glial cell culture Acute microglia isolation Mouse brain 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Annette Masuch
    • 1
  • Knut Biber
    • 2
  1. 1.Institute of Clinical Chemistry and Laboratory MedicineUniversity Medicine GreifswaldGreifswaldGermany
  2. 2.Department of Psychiatry and Psychotherapy, University Hospital FreiburgUniversity of FreiburgFreiburgGermany

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