A novel FACS-based method to isolate enriched populations of neurons, astrocytes, and microglia

We developed a novel FACS-based method to isolate enriched populations of neurons, astrocytes, and microglia. Whole mouse brains were digested, and populations of neurons, astrocytes, and microglia were subsequently enriched by FACS using a combination of positive gating on cell-type specific markers and stringent negative selection against markers of oligodendrocytes and endothelial cells. We confirmed a strong enrichment of corresponding cell-type specific genes in isolated populations and a reduction in gene markers of endothelial cells and oligodendrocytes, demonstrating that highly pure populations of neurons, astrocytes, and microglia were obtained. CNS cells from untreated wildtype mice were isolated using fluorescent activated cell sorting.