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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 effect of arsenious acid on differentiation of bone marrow stromal stem cells (MSCs) into neurons in vitro. Methods AMSCs were used to induce the morphological changes of human embryonic and adult mouse MSCs cultured in vitro. The effects of anti-Nestin, anti-MAP2, anti-βⅢtubulin, antiphospholipid basic protein (MBP) and glial fibrillary acidic Immunocytochemical staining of GFAP antibody and RT-PCR were used to detect the mRNA expression. 2-mercaptoethanol (BME) was used as a control. Results MSCs induced morphological features of neurons after induced by arsenite and BME. Arsenic trioxide induced cell morphology changes than the BME induction group appeared later, and the mortality rate is much lower than the latter. Immunocytochemistry showed Nestin, MAP2 and βⅢtubulin positive expression, MBP and GFAP negative expression. RTPCR visible Nestin, MAP2, βactin and β Ⅲ tubulin mRNA expression. Conclusion Both arsenite and BME can induce the differentiation of MSCs from human embryos and adult mice into neurons in vitro, but the induction of arsenite is even more mild.