Isolation and Characterization of Neuronal and Microglial Populations from Aging Mouse Brains

Adult neurons and microglia were isolated from the mice brain as previously described^{11 (link)}. In brief, the cortex of WT (1, 3, 5, or 9-months-old), APP/PS1 (1, 3, 5, or 9-months-old), APP/PS1 treated with N-AS (9-months-old) or APP/PS1 treated with aspirin (9-months-old) mice were minced in Hibernate A (Gibco)/B27 (Invitrogen) medium and dissociated using papain (Worthington) solution. After tissue trituration, cells were separated by Optiprep (Sigma-Aldrich) density gradient centrifugation. The purity of the fractionated neurons and microglia were acutely confirmed by neuronal (NeuN, mouse, 1:200, Millipore, MAB377) and microglial (Iba1, rabbit, 1:1000, Wako, 019-19941) markers and the isolated neurons and microglia were acutely analyzed for the acetylation assay and lipid mediators metabololipidomics.

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Lee J.Y., Han S.H., Park M.H., Song I.S., Choi M.K., Yu E., Park C.M., Kim H.J., Kim S.H., Schuchman E.H., Jin H.K, & Bae J.S. (2020). N-AS-triggered SPMs are direct regulators of microglia in a model of Alzheimer’s disease. Nature Communications, 11, 2358.

Publication 2020

Hibernate A papain Optiprep NeuN MAB377

Acetylation Adult Aspirin Assay Brain Cells Cortex Density gradient centrifugation Lipid Microglia Mouse Neurons Papain Rabbit Tissue

Corresponding Organization :

Other organizations : Kyungpook National University, Dankook University, Ulsan National Institute of Science and Technology, Hanyang University, Icahn School of Medicine at Mount Sinai

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Variable analysis

independent variables

Age of mice (1, 3, 5, or 9 months)
Genotype of mice (WT or APP/PS1)
Treatment of APP/PS1 mice (N-AS or aspirin)

dependent variables

Acetylation assay of isolated neurons and microglia
Lipid mediators metabololipidomics of isolated neurons and microglia

control variables

Tissue extraction (cortex)
Tissue dissociation and cell separation method (mincing, papain solution, Optiprep density gradient centrifugation)
Confirmation of cell purity (NeuN for neurons, Iba1 for microglia)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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