Protocol for metabolite extraction and sample preparation from primary astrocytes for mass spectrometry-based metabolomic analysis

Astrocytes play essential roles in supporting neuronal function, particularly through the regulation of brain energy metabolism. In response to physiological and pathological stimuli, astrocytes dynamically adjust their metabolic pathways and energy output. Here, we present a protocol for metabolite extraction and sample preparation from primary astrocytes for mass spectrometry analysis. We describe steps for integrating astrocyte culture and liquid chromatography-mass spectrometry (LC-MS) metabolite analysis to enable reproducible profiling of astrocytic energy metabolism under different experimental conditions. For complete details on the use and execution of this protocol, please refer to Chang et al.1.