Primary Central Nervous System (CNS) cell-based assays are vital tools in neuroscience research, utilizing neurons and glial cells directly isolated from brain tissue. These assays provide a more physiologically relevant model compared to immortalized cell lines, allowing researchers to study neuronal function, neurotoxicity, and disease mechanisms accurately. They are instrumental in drug discovery processes, enabling the assessment of drug effects on primary neurons for conditions like Alzheimer’s, Parkinson’s, and multiple sclerosis. Advancements in primary CNS cell-based assays enhance our understanding of the nervous system and accelerate the development of effective therapies.