Affiliation:
1. Department of Neurological Sciences , Rush University Medical Center , Chicago , USA
2. Division of Research and Development , Jesse Brown Veterans Affairs Medical Center , Chicago , USA
Abstract
Abstract
Objectives
Although microglia are activated in adult and aged brains resulting in neurodegenerative and neuroinflammatory disorders, most of the cell culture studies on microglia deal with neonatal microglia because of ease of isolation. Microglia could be isolated from adult brains, but it requires separation by density gradient centrifugation, magnetic beads, etc. Here, we describe a simple protocol of isolating highly purified microglia from adult mouse brains.
Methods
Our protocol involves dilution with sterile PBS or media, regular centrifugation, and plating on poly-D-lysine-coated flasks.
Results
These adult microglia expressed the inducible nitric oxide synthase in response to preformed α-syn fibril, an etiological reagent of Parkinson’s disease, and bacterial lipopolysaccharides, one of the prototype proinflammatory stimuli. Moreover, these adult microglia exhibited phagocytosis, which was stimulated by LPS treatment.
Conclusions
These results suggest that adult microglia isolated by our procedure are functional and that these adult microglia could be used for studies related to neurodegenerative disorders.
Funder
Center for Scientific Review
National Center for PTSD, U.S. Department of Veterans Affairs