Abstract:

BACKGROUND: During neural stem cell transplantation in the treatment of Parkinson’s disease, number of transplanted cells and differentiation ratio of dopaminergic neurons must be resolved. Effective in vitro proliferation of neural stem cells and large amount of directed differentiation of dopaminergic neurons are the key to solve above-mentioned problems.
OBJECTIVE: To investigate the differentiation of neural stem cells into dopaminergic neurons during induction of glial cell line-derived neurotrophic factor (GDNF) and interleukin-1β in vitro.
METHODS: Mesencephalon was isolated from mouse embryo ventral part following 12 days of pregnancy, and made into single cell suspension using trypsin digestion and mechanical trituration. Following centrifugation, neurospheres were subcultured by the mechanical method for 5-7 days, and then divided into groups to induce for 10-12 days. When 80% cells migrated from neurospheres differentiated into single cells, immunocytochemical identification and flow cytometry detection were utilized to determine the rate of tyrosine hydroxylase positive cells. 
RESULTS AND CONCLUSION: Neurospheres expressed nestin antigen, could differentiate into neuron specific enolase and glial fibral acidic protein-positive cells. Glial cell line-derived neurotrophic factor (GDNF) and interleukin-1β could significantly elevate the proportion of the differentiation of mesencephalic neural stem cells into tyrosine hydroxylase-positive neurons. The proportion was higher in the GDNF group, interleukin-1β group and GDNF + interleukin-1β group compared with the blank control group. In particular, the effect of GDNF + interleukin-1β was more significant. Results indicated that GDNF and interleukin-1β can obviously promote the differentiation of mesencephalic neural stem cells into enough number, mature morphology and mature function of dopamine neurons.