The research on the differentiation of dopaminergic neuron from neural stem cell induced by ascorbic acid and glial cell line-derived neurotrophic factor

Objective To investigate the effects of ascorbic acid(AA)and glial cell line-derived neurotrophic factor(GDNF)on the differentiation of neural stem cells into dopaminergic(DA)neurons.Methods To isolate and culture the neural stem cells(NSCs)from whole brain of neonatal Sprague Dawley rats(within 24h after birth),and identify them.AA or/and GDNF were added into the P 2 NSCs to induce DA neurons.After these cells were cultured for 10 days,the specific antigens of DA neuron including tyrosine hydroxylase(TH)and dopamine transporter(DAT)were detected by immunocytochemistry,and TH mRNA was detected by RT-PCR.Results Each group could be detected the expression of TH mRNA.The groups of 100μmol/L AA,10ng/ml GDNF and 100μmol/L AA+10ng/ml GDNF could increase the proportion of NSCs differentiated into TH-positive cells compared with the control group(P0.05).When treated with 100μmol/L AA+10ng/ml GDNF,the proportion of NSCs differentiated into TH-positive cells was significantly higher than 100μmol/L AA or 10ng/ml GDNF group alone(P0.05).Conclusion AA and GDNF could promote the differentiation of NSCs into DA neurons,when treated with GDNF+AA,the effect was stronger than AA or GDNF group alone.