大鼠胚胎中脑神经前体细胞体外诱导多巴胺能神经元的实验研究

Rudimental study of differentiation of neural precursor cells from mesencephalon of rat embryo into dopaminergic neurons

目的在体外多种条件下诱导大鼠胚胎中脑神经前体细胞,比较各自分化出多巴胺能神经元的比例。方法体外分离、培养出大鼠胚胎中脑腹侧神经前体细胞并传代,通过白介素-1α(IL-1α)、白细胞抑制因子(LIF)、胶质细胞衍生物神经营养因子(GDNF)、纹状体条件培养液和中脑膜裂解碎片诱导分化后,比较子代中酪胺酸羟化酶(TH)免疫阳性细胞的数量和形态变化。结果IL-1α能增加中脑神经前体细胞分化出的TH细胞数量,但形态不成熟;其他条件不能增加TH细胞数,但与IL-α共同作用时可使TH免疫阳性细胞形态变得成熟。结论IL-1α可以诱导中脑神经前体细胞分化多巴胺能神经元,LIF、GDNF、纹状体条件培养液和中脑膜裂解碎片在多巴胺能神经元的形态发育中起重要作用。

Objective To investigate some conditions which could in vitro induce neural precursor cells (NPCs) from mesencephalon of rat embryo to differentiate into dopaminergic neurons with a higher percentage. Methods NPCs were isolated from ventral mesencephalon of rat embryo, cultured and passaged in vitro, and then co-cultured with IL-1α, GDNF, LIF, striatal culture-conditioned media and mesencephalic membrane fragments. The number and morphology of TH immunopositive progenies were then examined. Results TH cells differentiated from mesencephalic NPCs co-cultured with IL-1α increased in number, but were immature in morphology. The other conditions co-cultured with mesencephalic NPCs could not increase the number of TH progenies, but could make TH cells more mature in morphology when they were added with IL-1α. Conclusion IL-1α could induce mesencephalic NPCs to differentiate into DA neurons, and LIF, GDNF, striatal culture-conditioned media and mesencephalic membrane fragments played an indispensable role in the morphogenesis of DA neurons.