The objective of this study was to show whether an in vitro model for Phenytoin-related cytoskeletal impairment could be helpful to investigate cerebellar side effects of Phenytoin (DPH). DPH dose-and time-dependently resulted in decreasing numbers of vital cells. Cells formed only a rarefield intercellular neuritic network. This effect was already evident 24 h after plating. Western-blot analysis revealed that the expression of the dendritic marker microtubule-associated protein 2 (MAP2) was dramatically decreased in DPH-treated cultures.