FETAL PORCINE VENTRAL MESENCEPHALON GRAFTS - DISSECTION PROCEDURE ANDCELLULAR CHARACTERIZATION IN CULTURE

The objective of this study was to develop an optimal dissection proce dure for fetal porcine ventral mesencephalon (VM) grafts and to charac terize the cellular composition of such an explant, in particular with respect to the dopaminergic and GABAergic components. We have used a monolayer cell culture system to study and identify the various VM cel l types. The in vitro development of the fetal VM cells and the effect of the addition of brain-derived neurotrophic factor (BDNF) was inves tigated during a culture period of 5 days. Extracellular dopamine leve ls were measured by means of high performance liquid chromatography (H PLC) with electrochemical detection (LCEC). Our results indicate that the ratio of dopaminergic to GABAergic neurons changed in favour of th e dopaminergic component when a more selective dissection technique wa s used. Although addition of BDNF to the cultures appeared to exert tr ophic influences on all the cellular components of pig fetal VM, this effect was most pronounced on the TH-positive cells, Highest extracell ular DA levels were found in the VM culture with the addition of BDNF and when a more selective dissection method was used. Our in vitro fin dings suggest that porcine fetal dopaminergic cells retain their poten tial for development and outgrowth after proper explantation and disso ciation. Anticipating on the results of ongoing transplantation studie s in rat, they suggest that pig fetal VM can be a suitable alternative for the use of fetal human VM as a graft for Parkinson's disease.