title: Microglia replenished OHSC: A culture system to study in vivo like adult microglia.
abstract: Recent data suggest that ramified microglia fulfil various tasks in the brain. However, to investigate this unique cell type cultured primary microglia are only a poor model. We here describe a method to deplete and repopulate organotypic hippocampal slice cultures (OHSC) with ramified microglia isolated from adult mouse brain creating microglia-replenished OHSC (Mrep-OHSC). Replenished microglia integrate into the tissue and ramify to a degree indistinguishable from their counterparts in the mouse brain. Moreover, wild-type slices replenished with microglia from TNFα-deficient animals provide similar results as OHSC prepared from microglia-specific TNFα-knockout mice (CX3CR1(cre) /TNFα(fl/fl) ). Furthermore, this study demonstrates that replenished microglia in OHSC maintain original functions and properties acquired in vivo. Microglia from ERCC1(Δ/ko) mice, a mouse model of accelerated aging, maintain enhanced Mac2 expression and their activated phenotype after replenishment to wild-type OHSC tissue. Thus, the present study demonstrates that Mrep-OHSC are a unique tool to construct chimeric brain slices allowing studying the function of different phenotypes of in vivo like microglia in a tissue culture setting. GLIA 2016 GLIA 2016;64:1285-1297.
authors.affiliation: Department of Psychiatry and Psychotherapy, University Hospital, University of Freiburg, Freiburg, Germany.
authors.forename: Annette
authors.initials: A
authors.lastname: Masuch
authors.affiliation: Department of Neuroscience, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Rianne
authors.initials: R
authors.lastname: van der Pijl
authors.affiliation: Department of Psychiatry and Psychotherapy, University Hospital, University of Freiburg, Freiburg, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: Füner
authors.affiliation: Department of Immunology, Weizmann Institute, Rehovot, Israel.
authors.forename: Yochai
authors.initials: Y
authors.lastname: Wolf
authors.affiliation: Department of Neuroscience, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Bart
authors.initials: B
authors.lastname: Eggen
authors.affiliation: Department of Neuroscience, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: Boddeke
authors.affiliation: Department of Psychiatry and Psychotherapy, University Hospital, University of Freiburg, Freiburg, Germany.|Department of Neuroscience, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Knut
authors.initials: K
authors.lastname: Biber
mesh_terms: D000818:Animals; D000073883:CX3C Chemokine Receptor 1 / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D037502:Galectin 3 / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000502:physiology*; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D017628:Microglia / Q000166:cytology / Q000502:physiology*; D018613:Microscopy, Confocal; D000066829:Neuroprotection / Q000502:physiology; D046509:Tissue Culture Techniques; D014409:Tumor Necrosis Factor-alpha / Q000172:deficiency / Q000235:genetics
chemical_list: D000073883:CX3C Chemokine Receptor 1; C471885:Cx3cr1 protein, mouse; D004268:DNA-Binding Proteins; D037502:Galectin 3; D014409:Tumor Necrosis Factor-alpha; D004720:Endonucleases; C499360:Ercc1 protein, mouse
keywords: ERCC1; NMDA; TNFα; hippocampus; neuroprotection; priming