THE DIFFERENTIATION OF HUMAN NEURAL STEM CELLS INTO DOPAMINERGIC NEURONS INDUCED BY IL-1?AND COMBINATION OF IL-1?,IL-11,LIF AND GDNF / 解剖学报

ABSTRACT

Objective To evaluate the effects of IL-1? and combination of IL-1?,IL-11,LIF and GDNF on inducing human neural stem cells(hNSCs)to differentiate into dopaminergic(DA)neurons in vitro. Methods A great deal of neurospheres was obtained by the technology of serum-free culturing and single-cell cloning,and was splitted into 3 groups,which were cultured in different media.In control group,the differentiation medium used only contained 10%FBS.In IL-1? group,the medium contained IL-1? and 10%FBS.In united factors group,the medium contained IL-1?,10%FBS supplemental with IL-11,LIF and GDNF.After 3 weeks,the tyrosine hydroxylase(TH)positive cells were detected by using immunofluorescence,and image processing about the number,the induced differentiational rate,the cell bodies' areas and the cells' perimeters of TH positive neurons was carried out.TH/MAP-2 double-immunofluorescence labeling was used to calculate the percentage of DA neurons in total neurons. Results In the control group,there were few TH positive neurons with poorly developed morphology.The presence of IL-1? induced more DA neurons,but these cells were still immature.In the united factors group,the number of maturer TH positive DA neurons was the most. Conclusion IL-1? has an obvious effect on inducing hNSCs derived from human fetal mescenphalon to DA neurons.The utilization of IL-1?,IL-11,LIF and GDNF in combination has a cooperative effect on inducing differentiation of hNSCs to mature DA neurons.