Protocol for sex-specific isolation of microglia, oligodendrocyte precursor cells, and astrocytes from neonatal rats using shake-off technique

Disease-related glial changes often show sex-specific differences, yet protocols for sex-specific isolation are limited. Here, we present a protocol for sex-specific isolation of microglia, oligodendrocyte precursor cells (OPCs), and astrocytes from neonatal rats using the shake-off technique. We describe steps for determining pup sex at post-natal days 1-2 and for dissecting, dissociating, and plating cortex cells. We then detail isolation procedures for each cell type. Downstream applications include immunocytochemistry, quantitative PCR, and RNA sequencing to assess cell-specific functions.