摘要
目的:探索大鼠骨髓间充质干细胞(mesenchymal stem cell,MSCs)在体外定向分化为神经元样细胞的方法并研究其死亡的影响因素。方法:分离纯化大鼠骨髓间充质干细胞并用2-巯基乙醇诱导分化为神经元样细胞,使用免疫组化方法鉴定其神经元特异性巢蛋白(Nestin)的表达,通过MTT、免疫组化及形态学方法观察丝裂原活化蛋白激酶激酶1/2(mitogen-activated protein kinase kinase 1/2,MEK1/2)阻断剂U0126及缩胆囊肽(cholecystokinin,CCK)对细胞转化及活性的影响。结果:诱导后MSCs分化为具有典型的神经元形态的细胞,免疫组化实验显示Nestin呈强阳性反应,同时免疫组化及MTT方法显示U0126可以显著提高细胞Nestin的表达强度和神经元样细胞的活性,CCK虽可轻度增强神经元样细胞的活性,且CCK可延长已分化神经元的存活时间,但其作用不具统计学意义。结论:大鼠MSCs在体外可定向诱导分化为神经元样细胞,且通过阻断MEK1/2信号通路可以提高其阳性分化率和细胞活性,提示大鼠MSCs诱导分化的神经元样细胞的死亡机制可能涉及MEK1/2信号通路。CCK可能具有对已分化神经元的保护作用。
Objective：To investigate the process of differentiation of adult rat mesenchymal stem cells into neuron-like cells in vitro and the following death.Methods：MSCs were isolated from adult rat bone marrow by density gradient and then expanded in medium as undifferentiated cells for 3-5 generations.The MEK1/2 inhibitor U0126 and CCK were introduced into the process of differetiation respectively to study their effects. MTF assay and immunocytochemistry were adopted to investigate the activity of neuron-like cells and the expression level of Nestin.Results：Most MSCs were induced to differentiate into neuron-like cells,high level of Nestin was expressed. And the MEK1/2 inhibitor U0126 apparently increased both the activity of neuron-like cells and the level of Nestin expression. While the effects of CCK in this process were negligible. Conclusion：MSCs could be induced in vitro to differentiate into neuron-like cells with high positive rate of differentiation and the MAPK signal pathway might have participated the death process of such neuron-like cells.
出处
《泸州医学院学报》
2007年第4期255-258,共4页
Journal of Luzhou Medical College
关键词
骨髓间充质干细胞
神经分化
缩胆囊肽
信号通路
细胞死亡
Mesenchymal stem cells
Neuron differentiation
Cholecystokinin
Signal pathway
Cell death
