Protocol for assessing molecular signatures and metabolic function of primary murine cortical astrocytes

Astrocytes are the most numerous brain cells and are central to the regulation of brain metabolism. Here, we present a protocol for identifying molecular signatures and assessing the metabolic function of primary murine cortical astrocytes. We describe steps for extracting, isolating and culturing astrocytes, characterizing these cells with immunocytochemistry, and extracting their metabolites. We then detail procedures for assessing mitochondrial function with challenges, RNA extraction for bulk sequencing, and DNA extraction for sequencing and epigenetics.