摘要
目的探讨细胞因子白介素-1α(IL-1α)及与白介素-11(IL-11)、白血病抑制因子(LIF)和胶质细胞源性神经生长因子(GDNF)联合应用对体外诱导人神经干细胞定向分化为多巴胺神经元的影响。方法采用无血清培养技术和单细胞克隆技术、免疫荧光双标技术。结果对照组仅有极少量的神经干细胞分化为不成熟的多巴胺神经元;IL-1α组分化的多巴胺神经元较多,但细胞形态欠成熟;联合因子组分化的多巴胺神经元最多,细胞较为成熟。结论IL-1α具有诱导人神经干细胞向多巴胺神经元分化的作用,联合应用IL-1α、IL-11、LIF和GDNF对人神经干细胞向成熟的多巴胺神经元分化具有协同作用。
Objective To evaluate the effects of lL-1α and combination of IL-1α, 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-1α group,the medium contained IL- 1α and 10 % FBS. In united factors group, the medium contained IL- 1 α, 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-mmunofluorescence 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 lL-1α 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 lL-1α has an obvious effect on inducing hNSCs derived from human fetal mescenphalon to DA neurons. The utilization of lL-1α, IL-11, LIF and GDNF in combination has a cooperative effect on inducing differentiation of hNSCs to mature DA neurons.
出处
《解剖学报》
CAS
CSCD
北大核心
2005年第4期351-355,共5页
Acta Anatomica Sinica
基金
江苏省神经再生重点实验室科研基金资助项目(No.200201)
