胞磷胆碱体外诱导人脐带间充质干细胞分化为神经元样细胞的实验研究

Experimental Study on Effect of Citicoline in Inducing Differentiation of Human Umbilical Cord-derived Mesenchymal Stem Cells to Neuron-like Cells

目的探讨胞磷胆碱(CDPC)作为神经保护剂在B-巯基乙醇诱导人脐带间充质干细胞(hUMSCs)向神经元样细胞的诱导分化中的作用。方法分别用胞磷胆碱(CDPC,A组)、B-巯基乙醇(B-ME,B组)、胞磷胆碱联合B-巯基乙醇(C组)、生理盐水(D组)体外诱导第3代hUMSCs向神经元样细胞分化,免疫荧光技术鉴定分化细胞巢蛋白(Nestin),比较各组Nestin阳性细胞百分率。结果 A组与D组比较Nestin阳性细胞百分率无统计学差异,B组诱导1、10 h后阳性率高于C组,但诱导24 h后细胞大片死亡,无法检测阳性率。C组诱导24 h后则未出现细胞大片死亡现象。结论 CDPC在体外不能诱导hUMSCs分化成神经元样细胞,但与B-ME联用后可以降低B-ME的细胞毒性,保护神经细胞,为CDPC在中枢神经系统疾病干细胞治疗的临床应用提供了理论依据。

Objective To investigate the effect of citicoline （CDPC）, as a neuroprotective agent, in Bmercaptoethanol inducing the differentiation of human umbilical cord-derived mesenchymal stem cells （hUMSCs） cells into neuron-like cells. Methods The 3rd generation hUMSCs were induced differentiate into neuron-like cells by Citicoline （ CDPC, group A）, B-mercaptoethanol （ B-ME, group B）, Citicoline joint B-mercaptoethanol （group C）, Saline （ group D） in vitro. The expressions of Nestin were identified by immunofluorescence techniques, and the rate of the Nestin positive staining was compared. Results There was no significant difference between group A and group D in the rate of positive staining cells. The Nestin ＋ cells level of group B were dramatically higher than group C （ P 〈 0.01 ） after 1 h and l0 h treatment, most of the cells were dead after 24 h. Cell apoptosis did not appear obviously in group C. Conclusion CDPC can not induce hUMSCs differentiate into neuron-like cells in vitro, but can protect the nerve cells by reducing the cytotoxicity of B-ME, which provides a theoretic evidence on the clinical application of CDPC in the treatment of central nervous system diseases with stem cell therapy.