Grafting of Dopaminergic Ventral Mesencephalic Slice Cultures to the Striatum of Adult Rats

Abstract Live storage of dopaminergic neurons before intracerebral grafting will allow pregrafting examination and manipulation of the cells, as well as pooling and mixing of cells from several donors. In this study we examined whether mesencephalic dopaminergic neurons, grown in organotypic cultures for 1 week, would survive subsequent grafting to the adult rat striatum. Slices of ventral mesencephalon from neonatal rats were grown by the Roller drum method for 1 week and then grafted into the striatum of adult rats, with and without preceding 6-hydroxydopamine lesions of their nigrostriatal pathway. Using immunocytochemical staining for tyrosine hydroxylase, cultured dopaminergic neurons were found to survive and to extend fibers into the host striatum when examined 4, 7, 14, 28, and 87 days after grafting. When compared with slices of noncultured mesencephalic tissue from 1-week-old rats the slice culture period did not significantly reduce the number of surviving tyrosine hydroxylase positive neurons. From this we conclude that slice cultures can be used for transient storage of dopaminergic donor tissue before intracerebral grafting. The surviving tyrosine hydroxylase positive neurons in control grafts from 1-week-old rats, furthermore, extends the time frame of possible donor ages used for grafting.