Primary Rat Mesencephalic Neuron-Glia, Neuron-Enriched, Microglia-Enriched, and Astroglia-Enriched Cultures

Degeneration of dopaminergic neurons in the substantia nigra is a hallmark of Parkinson’s disease (PD). However, despite decades of research, the cause of PD and the underlying mechanism of action responsible for the progressive degeneration of nigral dopaminergic neurons remain poorly understood (1 ). The creation of rodent and primate models for PD has provided a valuable tool in the study of the pathogenetic progression of the disease (2 ). On the other hand, primary neural cell cultures have become extremely valuable in the delineation of the molecular and cellular mechanisms of neuronal death. In light of the increasing appreciation of the role brain immune cells play in the neurodegenerative process (3 ), establishment of enriched primary cultures of neurons, microglia, and astroglia enables the dissection of the complex in vivo system in an in vitro setting. The utility of these cultures has helped gain critical information on the role each cell type plays and the potential factors produced by individual cell types that contribute to the neurodegeneration (4 –10 ). Here we describe our laboratory’s routinely used procedures for establishing primary mixed mesencephalic neuron-glia, mixed glia, neuron-enriched, microglia-enriched, and astroglia-enriched cultures.