Chitosan is a popular biomaterial used in tissue engineering. Fibers of chitosan could provide a favorable anatomical substrate for cell growth which provides a promising application for axonal regeneration during peripheral injury. Neuroepithelial stem cells (NEPs) are the most primitive neural stem cells with multipotential for neuronal and glia differentiation. To assess the biocompatibility between NEPs and chitosan fibers, and to explore whether the NEPs have the ability to differentiation on chitosan fibers, NEPs were harvested from the neural tube and seeded on chitosan fibers in in vitro culture. The biocompatibility of chitosan fibers was tested by MTT assays. The growth and survival were observed by light and scanning electronic microscope at different times in culture. And, the differentiation of NEPs was examined by immunocytochemical staining. The results indicated that NEPs could grow on the chitosan fibers and attach firmly to the surface of fibers. On chitosan fibers, NEPs could differentiate into neurons and glia. Our study demonstrated that chitasan fibers had a good biocompatibility with NEPs which affords a potential alternative for the repair of peripheral nerve injury.