Beta-catenin is a key player in the canonical beta-catenin mediated Wnt-pathway during development of many cell types. It has a major role in activation of TCF-dependent transcription. Prerequisite of gene activation is its translocation into the nucleus. In this study the human neural progenitor cell line ReNcell VM 197 (ReNeuron, Guildford, UK) derived from 10 weeks old fetal ventral mesencephalon was used to investigate the influences of the Wnt-pathway on beta-catenin translocation. These cells have the capacity to differentiate into different glial and neuronal cell types, especially of the dopaminergic phenotype. Here we show that they differentiate under the control of beta-catenin. beta-catenin signals were quantified in different cellular compartments during the early differentiation phase of the cells by both quantitative image analysis of confocal image stacks and subcellular fractionation followed by Western blotting. To investigate the influence of Wnt signals and Wnt pathway components on the beta-catenin localization, the cells were treated with either the glycogensynthase kinase (GSK)- 3beta inhibitor SB216763, or WIF-I (Wnt inhibitory factor-I), an inhibitor of free Wnt proteins. Additionally Wnt3a, a stimulator of the canonical Wnt-pathway, was tested to investigate the upstream effect of this pathway. The results show that the nuclear beta-catenin concentration increases after induction of cell differentiation by growth factor removal. Treatment with SB216763 enhanced this effect, whereas WIF-I treatment suppressed it. Furthermore, treatment with SB216763 resulted in a significant enhancement of the neuronal, especially the dopaminergic phenotype as detected by an increase of cells expressing beta-III tubulin and tyrosine hydroxylase, respectively. After treatment with WIF-I a concentration-dependent decrease of cells with neuronal phenotype occurred. After Wnt-3a treatment Western blot analysis revealed accumulation of nonphospho-ß-catenin and of cytoplasmic ß-catenin which is accompanied by phosphorylation of LRP6 and Dvl. These results show a beta-catenin dependent enhancement of the differentiation of neural progenitor cells into the neuronal, especially dopaminergic, phenotype and thus might have relevance for approaches of cell replacement therapy of Parkinson’s disease.