Development and survival of rat embryonic mesencephalic dopaminergic neurones in serum-free, antioxidant-rich primary cultures.

Mesencephalic dopaminergic neurones typically require the presence of serum for their survival in culture. However, the present study, outlines how neurones from the rat ventral mesencephalon (E14-16) were successfully cultured in serum-free, antioxidant-rich Neurobasal medium supplemented with B27 components. Moreover, immunostaining with mouse monoclonal microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) from day 1 to 7 in vitro revealed these cultures were primarily neuronal (80-95%). Additionally, immunostaining with tyrosine hydroxylase (TH) revealed these cultures contained a relatively constant population of TH-positive neurones (5%) which were presumed to be dopaminergic. These primary cultures offer considerable advantages for the study of mesencephalic, TH-positive, dopaminergic neurones under conditions where milieu can be readily manipulated in the virtual absence of glia and without the confounding influence of serum.