Recent medical cell therapy using polymeric biomaterial-loaded stem cells with the capability of differentiation to specific neural population has directed focuses toward the recovery of central nervous system. Definite culture conditions that regulate cell morphogenesis, proliferation, survival, and differentiation to lineage specific cells have a great influence on the cells' life. In this research, random and aligned poly(lactic-co-glycolic acid) fibrous scaffolds with nano-structure were prepared, and the effect of nano-fiber morphology on the pluripotent P19 embryonic carcinoma cell proliferation and neural differentiation in response to retinoic acid induction was studied. Fiber morphology was assessed by scanning electron microscope (SEM). For in vitro studies, SEM, (4,5-dimethylazol-2-yl)-2,5-diphenyl-2H tetrazolium bromide assay, and immunofluorescent staining were carried out for the assessment of biological properties of the seeded cells. Our results showed that, because of their similarity with natural extracellular matrix, the aligned fibrous substrates provide the suitable surface for cell function as well as the augmentation of cell differentiation. Copyright (c) 2014 John Wiley & Sons, Ltd.