Mesenchymal stem cells have been extensively used for cell-based therapies especially in neuronal diseases. Studies still continue to delineate mechanisms involved in differentiating mesenchymal stem cells into neuronal cells under experimental conditions as they have low mortality rate and hence, the number of cells available for experiments is much more limited. Culturing and differentiating of neuronal cell is more challenging as they do not undergo cell division thus, bringing them to differentiate proves to be a difficult task. Here, the aim of this study is to investigate whether Juglans regia L. (walnut oil) differentiates multipotent, C3H10T1/2 cells, a murine mesenchymal stem cell line, into neuronal cells. A simple treatment protocol induced C3H10T1/2 cells to exhibit a neuronal phenotype. With this optimal differentiation protocol, almost all cells exhibited neuronal morphology. The cell bodies extended long processes. C3H10T1/2 cells were plated and treated with walnut oil post 24 h of plating. The treatment was given (with walnut oil treated cultures with or without control cultures) at different concentrations. The cultured cells were then stained with cresyl violet acetate solution which was used to stain the Nissl substance in the cytoplasm of the induced neuronal culture. The results indicated that the C3H10T1/2 cells differentiated into neuronal-like cells with long outgrowths of axon-like structures able to take up the cresyl violet acetate stain indicating their preliminary differentiation into neuronal-like morphology with walnut oil treatment. Treating the mesenchymal stem cells can in future establish a cultured mesenchymal stem cell line as neuronal differentiating cell line model.