亚砷酸诱导骨髓基质干细胞向神经细胞的分化作用

Neuronal development of bone marrow stromal stem cells induced by arsenious acid

目的研究亚砷酸体外诱导骨髓基质干细胞(MSCs)向神经细胞分化的作用。方法应用亚砷酸诱导体外培养的人胚胎和成年小鼠MSCs,观察诱导过程中的形态学变化;应用抗Nestin、抗MAP2、抗βⅢtubulin、抗磷脂碱性蛋白(MBP)及抗胶质纤维酸性蛋白(GFAP)抗体进行免疫细胞化学染色,逆转录聚合酶链反应(RTPCR)检测mRNA表达,以2巯基乙醇(BME)作为对照。结果经亚砷酸和BME诱导后,MSCs均表现为神经元细胞的形态特征。亚砷酸诱导组细胞形态变化较BME诱导组出现晚,且死亡率远低于后者。免疫细胞化学示Nestin、MAP2和βⅢtubulin阳性表达,MBP及GFAP为阴性表达。RTPCR可见Nestin、MAP2、βactin和βⅢtubulinmRNA阳性表达。结论亚砷酸和BME均可在体外诱导人胚胎和成年小鼠MSCs向神经细胞分化,但亚砷酸诱导更为缓和。

Objective To investigate the regulation of neuronal development of human fetal and Kunming adult mouse marrow stromal stem cells （MSCs） induced by arsenious acid in vitro. Methods Morphological changes of neuronal development during the induction of MSCs caused by arsenious acid and β-mercaptoethanol were observed. Induced cells were stained immunocytochemically with Nestin, MAP2, βⅢ-tubulin, MBP and GFAP. RT-PCR was used to detect the mRNA and protein production. Resuits MSCs showed morphological changes after induction with arsenious acid and β-mercaptoethanol. Typical neuron-like morphology appeared gradually with the prolongation of time. But mortality rate of MSCs caused by β-mercaptoethanol was really higher than that by arsenious acid. These cells were positively stained with Nestin, MAP2 and βⅢ-tubulin antibodies, but negatively with MBP and GFAP. RT-PCR revealed Nestin, MAP2 and βⅢ-tubulin mRNA were positively expressed respectively. Conclusion Human fetal and adult mouse marrow stromal stem cells can develop into neural ceils by induction of arsenious acid and β-mercaptoethanol in vitro. Arsenious acid is better than β-mercaptoethanol in view of gradual induction and lower mortality rate.