ObjectiveTo establish a method of isolation and culture of neural stem cells(NSCs). MethodsTissues of ventral midbrain were isolated from a rat embryo,and the NSCs were cultured stimulated with basic fibroblast growth factor(bFGF)in vitro.The cells were identified by immunocytochemistry.ResultsNSCs proliferated into neurosphere in symmetric and non-symmetric cleavage ways,and differentiated into neuron, astroglia and oligodendrosyte. ConclusionsThe method has been established to isolate and culture the neural stem cells.

ObjectiveTo determine survival and differentiation of cultured neural stem cells (NSCs) into viable and functional neurons upon transplantation into the brain of MPTP-induced Parkinson's Disease (PD) mode micel.MethodsMice model of PD was established with intraperitoneal (i.p.) injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP,40mg/kg) twice interspersed by 16 hr. Tissues isolated from an embryonic rat midbrain were cultured in clonal density. After labeled with 5-bromo-2′-deoxyuridine (BrdU), the NSCs were transplanted into the uni- or bilateral striatum of PD mice. Tyrosine hydroxylase (TH) immunofluorescence was used to evaluate the toxicity of MPTP on the neural cells in the substantia nigra. Immunohistochemistry and laser confocal were used to detect the survival and differentiation of transplanted NSCs.ResultsTH positive neural cells were significantly reduced in the substantia nigra after MPTP injection. Immunohistology showed that the uni-or bilateral transplanted NSCs could survive in the brains of PD model mice.Some transplanted NSCs could properly differentiate into targeted TH positive neural cells in vivo.ConclusionsThe transplanted multipotent NSCs could survive, differentiate into functional dopamine neurons in the brains of PD model mice.

ObjectiveTo observe the effects of neural stem cells transplantation on the behavioral deficits of Parkinson's disease mice.MethodsSpontaneous movement,Morris Water Maze and Rotarod were adopted to evaluate the behavioral changes.ResultsCompared with the control,the time of spontaneous movement was decreased,the latency of Water Maze was lengthened,and the time of Rotarod was shortened in the Parkinson's disease(PD) mice. These deficits were improved 2,4 and 8 weeks after unilateral and bilateral transplantation of neural stem cells.ConclusionsThe transplantation of neural stem cells can improve the behavioral deficits in the PD mice.