title: An optimized protocol for the acute isolation of human microglia from autopsy brain samples.
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.
authors: Section Medical Physiology, Department of Neuroscience, UMCG-RuG, Groningen, The Netherlands.
authors: Marta
authors: M
authors: Olah
authors: Divya
authors: D
authors: Raj
authors: Nieske
authors: N
authors: Brouwer
authors: Alexander H
authors: AH
authors: De Haas
authors: Bart J L
authors: BJ
authors: Eggen
authors: Wilfred F A
authors: WF
authors: Den Dunnen
authors: Knut P H
authors: KP
authors: Biber
authors: Hendrikus W G M
authors: HW
authors: Boddeke
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D017304:Annexin A5 / Q000378:metabolism; D001253:Astrocytes / Q000145:classification / Q000502:physiology*; D001344:Autopsy / Q000379:methods; D001921:Brain / Q000166:cytology*; D002452:Cell Count; D002465:Cell Movement; D002499:Centrifugation, Density Gradient / Q000379:methods; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008297:Male; D008875:Middle Aged; D010587:Phagocytosis / Q000502:physiology; D011205:Povidone; D017382:Reactive Oxygen Species / Q000378:metabolism; D012822:Silicon Dioxide; D055815:Young Adult
chemical_list: D017304:Annexin A5; D017382:Reactive Oxygen Species; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone