Strategies of Schwann cell (SC) transplantation for regeneration of peripheral nerve injury involve many limitations. Stem cells can be used as alternative cell source for differentiation into Schwann cells. Given the high potential of neural crest-derived stem cells for the generation of multiple cell lineages, in this research, we considered whether olfactory ectomesenchymal stem cells (OE-MSCs) derived from neural crest can spontaneously differentiate into SC lineage. OE-MSCs were isolated from human nasal mucosa and characterized by the mesenchymal and neural crest markers. The cells were cultured in glial growth factors-free medium and further investigated in terms of the phenotypic and functional properties. Immunocytochemical staining and real-time PCR analysis indicated that the cultured OE-MSCs expressed SCs markers, SOX10, p75, S100, GFAP and MBP, differentiation indicative. It was found that the cells could secrete neurotrophic factors, including brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). Furthermore, after co-cultured with PC12, the mean neurite length was enhanced by OE-MSCs. The findings indicated that OE-MSCs could be differentiated spontaneously into SC-like phenotypes, suggesting their applications for transplantation in peripheral nerve injuries.Strategies of Schwann cell (SC) transplantation for regeneration of peripheral nerve injury involve many limitations. Stem cells can be used as alternative cell source for differentiation into Schwann cells. Given the high potential of neural crest-derived stem cells for the generation of multiple cell lineages, in this research, we considered whether olfactory ectomesenchymal stem cells (OE-MSCs) derived from neural crest can spontaneously differentiate into SC lineage. OE-MSCs were isolated from human nasal mucosa and characterized by the mesenchymal and neural crest markers. The cells were cultured in glial growth factors-free medium and further investigated in terms of the phenotypic and functional properties. Immunocytochemical staining and real-time PCR analysis indicated that the cultured OE-MSCs expressed SCs markers, SOX10, p75, S100, GFAP and MBP, differentiation indicative. It was found that the cells could secrete neurotrophic factors, including brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). Furthermore, after co-cultured with PC12, the mean neurite length was enhanced by OE-MSCs. The findings indicated that OE-MSCs could be differentiated spontaneously into SC-like phenotypes, suggesting their applications for transplantation in peripheral nerve injuries.

Numerous studies have shown the positive effects of rosmarinic acid on the nervous system. Rosmarinic acid as a herbal compound with anti-inflammatory effects can prevent the destructive effect of inflammation on the nervous system. Furthermore, various studies have emphasized the advantages of three dimensional (3D) culture over the two dimensional (2D) culture of cells. In this study, thermosensitive chitosan-based hydrogel as a 3D scaffold with the combination of chitosan (CH), beta-glycerol phosphate(βGP) and hydroxyl ethyl cellulose (HEC) CH-βGP-HEC loaded with rosmarinic acid was used to induce neuronal differentiation in human Wharton jelly stem cells. Also, cells were divided into eight groups in order to evaluate the effect of 3D cell culture and to compare gene expression in different induction conditions. The results of gene expression analysis showed the highest expression of neuronal markers in WJMSCs cultured in CH-βGP-HEC loaded with differentiation medium and rosmarinic acid. According to the results of gene expression, rosmarinic acid alone have positively effect on the induction of expression of neural markers. This positive effect is enhanced by cell culture in 3D conditions. This study shows that rosmarinic acid can be considered an inexpensive and available compound for use in neural tissue engineering. The results of this study indicate that rosmarinic acid can be considered a cheap and available compound for use in neural tissue engineering. The results of our study also emphasize the need to study stem cell differentiation under 3D culture conditions.