摘要
目的:研究化学方法体外诱导大鼠骨髓间充质干细胞向神经样细胞的可持续性分化。方法:体外培养大鼠骨髓间充质干细胞至P5代,流式细胞术检测细胞表面标志物,将细胞分为丁羟茴醚(BHA)诱导组、β巯基乙醇(BME)诱导组和对照组,分别进行化学诱导分化,通过荧光定量PCR、Western印迹和免疫细胞化学法分别检测巢蛋白、神经元特异性烯醇化酶(NSE)、胶质原纤维酸性蛋白(GFAP)等神经细胞标志分子的表达情况。结果:BHA诱导组高表达GFAP,而BME诱导组高表达巢蛋白,2组NSE表达均较弱;同时在诱导结束后继续培养的过程中,2组神经标志分子表达量持续降低。结论:2种方法都能诱导骨髓间充质干细胞向神经样细胞分化,但分化结局不同,BHA诱导分化后细胞主要表达星形胶质细胞表面标志GFAP,而BME诱导分化后细胞主要表达神经前体细胞表面标志巢蛋白;此外,通过化学方法诱导分化的效果是不可持续的,在诱导结束后继续培养的过程中神经标志分子表达逐渐下调。
Objective:To explore the sustainable differentiation of bone mesenchymal stem cells(BMSCs)into neurons in vitro.Methods:BMSCs passaged five times were used and divided into three groups:butylated hydroxyanisole(BHA)-induced group,β-mercaptoethanol(BME)-induced group and control group.The expression levels of nestin,neuron-specific enolase(NSE)and glial fibrillary acidic protein(GFAP)were detected through real-time PCR,Western blot and immunocytochemistry respectively.Results:After induced differentiation,GFAP expression was higher in BHA-indcuced group than that in BME-induced group,and nestin was higher in BME group than that in BHA group and these two genes were decreased during the next induction-free cultivation.NSE was not significantly changed in both induced groups.Conclusion:BHA-induced BMSCs differentiated into astrocytes with high expression of GFAP,while BME-induced BMSCs differentiated into neural precursor cells with high expression of nestin;and the differentiation effect of both chemical-induced methods are discontinued upon the induction is withdrawed.
作者
黄鑫
刘宇
卢宏
HUANG Xin;LIU Yu;LU Hong(Department of Neurology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
出处
《生物技术通讯》
CAS
2017年第6期793-797,共5页
Letters in Biotechnology
基金
河南省卫生厅中青年科技创新人才项目
关键词
骨髓间充质干细胞
神经细胞
诱导分化
bone mesenchymal stem cells(BMSCs)
neuron
induced differentiation
