Enhanced Neural Differentioation of Epidermal Neural Crest Stem Cell by Synergistic Effect of Lithium Carbonate and Crocin on BDNF and GDNF Expression as Neurotrophic Factors

Neurodegenerative diseases are incurable and debilitating conditions that result in progressive degeneration of nerve cells. Due to the complexity of conditions in neurodegenerative diseases, combination therapy including cell therapy and drug therapy is important as a new therapeutic strategy. Epidermal neural crest stem cells (EPI-NCSCs) are as one of the best option in cell therapy for various neurologicaldiseases. In this study, the effect of Lithium carbonate and Crocin, with considering of their effects on cellular signaling pathways and neuroprotective properties were investigated on expression of neurotrophic factors BDNF and GDNF in EPI-NCSCs. Materials and Methods:EPI-NCSCs were isolated from the Whisker hair follicles of the 2-week-old male Wistar rat and cultured in collagen-coated culture plates (1mg/ml). After EPINCSCs emigration, adhering cells removed by trypsin and subsequently subcultured. Then, after 72 hours oftreatment with different concentration of drugs [Lithium (0.1, 0.5, 1, 1.2, 1.4, 1.6, 1.8, 2, 4, 8 mM), Crocin (12.5, 50, 100, 200, 500, 1000, 1500, 2000 ,2500 μM) and lithium (1mM) + Crocin (12.5, 50, 100, 500, 1000, 1500, 2000 ,2500 μM)], and trial doses were selected by MTTassay. The cells were treated with selected concentration (Lithium 1 mM, Crocin 1.5 mM, Lithium 1mM and Crocin 1 mM) for 7 days. Results: The Real Time PCR results indicated an increasing in expression of BDNF and GDNF in treated cells as compared with control(*p<0.05, **p<0.01 and ***p<0.001). Conclusion: The results in this study confirmed and supported the neuroprotective/neurogenesis effects of Lithium and Crocin, it also showed that the proposed protocol can be used to increase EPI-NCSCs differentiation potential into neural cells in cell therapy and combination therapy of neurodegenerative diseases.