Experimental study of the functions of GDNF and dbcAMP on the induction of differentiation of neural stem cells into dopaminergic neurons

Objective To obtain a considerable number of dopaminergic cells through the use of GDNF and dbcAMP on rat embryonic neural stem cells for directed differentiation and therefore lay the foundation for the next cell transplantation. Methods Neural stem cells of 14-16 days pregnant Wistar rat were isolated and cultured,cells were subcultured to detect its self-renewal capacity and its Nestin and Brdu protein,identification,and then take use of rat neural stem cells that differentiation directed by GDNF and dbcAMP,the expression of TH positive cells were detected by immunocytochemistry and immunofluorescence methods. Results Cultured neural stem cells expressed Nestin and Brdu. A significant increase in the number of neurospheres after one week of in vitro culture were observed,the volume increased too. GDNF and dbcAMP could increase TH expression respectively and there were significant differences compared with the serum control group( P 0. 05). Conclusion In the role of the neurotrophic factor,neural stem cells can rapidly proliferate and form neurospheres. Both GDNF and dbcAMP can result in the induction of differentiation of neural stem cells into dopaminergic neurons,GDNF induces the highest performance in 20 μg /L and dbcAMP induces the highest performance in 200 μmol /L,both in a does-dependent manner. There is significant difference in TH cells positive rate between combination of induction and independent induction.