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.
Cheung
N S
NS
Department of Pharmacology, Monash University, Clayton, Victoria, Australia.
Hickling
Y M
YM
Beart
P M
PM
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002470:Cell Survival / Q000502:physiology*; D002478:Cells, Cultured; D004298:Dopamine / Q000378:metabolism*; D008636:Mesencephalon / Q000254:growth & development* / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000975:Antioxidants; D004298:Dopamine