Differentiation of humanmesenchymal stem cells (hMSC) to neural cells on Nano-scaffolds is a promising method for the treatment of the damaged nervous system through bionanomaterial-cell transplantation. The hMSC’s multipotential features have been discovered in various tissue engineering researches. This investigation shows the in-vitro development and neural differentiation of hMSC in 3D and 2D environments. The 3D environment which used in this study is nanofibrous polycaprolactone (PCL). The differentiation potential of mesenchymal stem cells (MSCs) to neural cells, on the random polycaprolactone (PCL) nanofibrous scaffolds, and tissue plate was examined. Researches have proved that interaction of extracellular nanofibrous matrix with in-vivo cells, gives mechanical maintenance to the cells and plays a functional role in the control of cellular behaviour. Stem cells are developing as a fundamental tool in the evolution of tissue engineering and regenerative medication. PCL characterization was determined employing scanning electron microscopy (SEM). Agents like, retinoic acid, epidermal growth factor (EGF), fibroblast growth factor (FGF-2), and Ibmx, which they are neural inducing agents, added in DMDM/F12 to differentiate MSCs to neural cells. Reproduction of mesenchymal cells on PCL nanofibrous scaffolds and neural morphology revealed through a scanning electron microscope (SEM) and optical microscope outcomes. The differentiated mesenchymal cells on nanofibrous scaffolds express neural gene markers including; - tubulin III and Map2 on the day of 14. Our investigation recommends the potential usage of differentiated neural cells from hMSCs on Nano-scaffolds toward the improvement of neural cells. This study conducted in 2011.