【摘要】 比较β-巯基乙醇(BME)和bFGF分别诱导大鼠骨髓基质干细胞(MSC)所分化神经元样细胞的生物学性状。BME和bFGF分别作用于大鼠的MSC,分别于9h和5d后,进行神经元样细胞的细胞计数,然后用免疫荧光进行神经元特异性蛋白NF200的鉴定,RT-PCR的方法分析神经元特异性基因NF的表达;western blotting对检测神经元特异性蛋白NF200的表达。BME诱导的细胞在加入药物1h后形态发生改变,9h时有突起伸出,胞体变亮;bFGF诱导的细胞在诱导后第3天开始有突起伸出,呈神经元样,第5天突起之间相互连接,第7天胞体死亡;而2种药物所分化的神经元样细胞NF200的免疫荧光均成阳性,但bFGF诱导的神经元样细胞的阳性率要显著高于BME所诱导的神经元样细胞;在RT-PCR中,bFGF诱导的神经元样细胞神经元特异性基因NF200的表达要高于BME所诱导的细胞;western blotting中,bFGF诱导的神经元样细胞神经元特异性蛋白NF200的表达要高于BME所诱导的细胞。bFGF和BME均可诱导MSC分化为神经元样细胞。但bFGF所诱导的神经元样细胞在从形态上更接近于神经元,而神经元特异性基因NF200mRNA及神经元特异性蛋白NF200蛋白的表达,均高于BME所诱导的细胞。更多还原

【Abstract】 To compare the biological characteristics of neuron-like cells from rat mesenchymal stem cells (MSC) induced by β-mercaptoethanol (BME) and bFGF respectively. BME and bFGF were added to rat MSC respectively for 9 h and 5 d. The neuron-like cells from MSC with the neuron specific protein NF200 were identified using immunofluorescence. The gene expression of NF were analyzed using RT-PCR and the protein expression of NF200 were analyzed using Western blotting. MSC induced by BME appeared morphology change in an hour and the neurite can be seen at six hour. Some cell bodies became lighter and died. MSC induced by bFGF had neurite in the third day, and appeared network structure in the fifth day, then cells died in the seventh day. At the same time, The result of immunofluorescence showed the NF200 expression of neuon-like cells induced by two ways were both positive, but MSC induced by bFGF were stronger than those induced by BME. The results of RT-PCR and western blotting were same as that of immunofluorescence. Both bFGF and BME can induce rat MSC to neuron-like cells, but the neuon-like cells induced by bFGF were more like real neurons in morphology and expression of neuron specific protein and mRNA.更多还原