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为了研究大鼠骨髓间充质干细胞(MSCs)诱导分化为神经细胞的表型特征,利用贴壁培养法获得骨髓MSCs。化学诱导剂二甲基亚砜(DMSO)和β-巯基乙醇(BME)联合诱导MSCs。免疫组织化学染色检测神经元特异性标志物MAP2、NSE和NF的表达,以及胶质细胞标志物GFAP的表达。硫堇-伊红染色检测细胞内Nissl体。结果表明,DMSO和BME联合诱导MSCs后48h,80%以上的细胞变为神经元样形态,胞体发出数个突起,有的似轴突,突起交织成网。免疫组化结果表明,诱导后细胞表达神经元特异性标志物MAP2、NSE和NF,其诱导率分别为88.6%,87.1%和85.8%。神经干细胞的特征性生物学标记nestin的诱导率在诱导后2h和10h较高,分别为48%和71.4%,而在诱导后48h仅为0.06%。诱导后细胞不表达GFAP。Nissl染色结果表明,诱导后细胞胞质中含有Nissl体。本研究结果证明DMSO和BME联合诱导MSCs可获得具有神经元表型特征的MSCs源性神经细胞。
To investigate the phenotypic characteristics of rat bone marrow mesenchymal stem cells (MSCs) differentiated into neurons, MSCs were obtained by adherent culture. Induction of MSCs with chemical inducer dimethyl sulfoxide (DMSO) and β-mercaptoethanol (BME). Immunohistochemical staining was used to detect the expression of neuron specific markers MAP2, NSE and NF, as well as glial cell marker GFAP. Thionine-eosin staining for intracellular Nissl body. The results showed that more than 80% of cells became neuron-like at 48h after MSCs were induced by DMSO and BME. The results of immunohistochemistry showed that the induction rates of neurons specific MAP2, NSE and NF were 88.6%, 87.1% and 85.8% respectively. The induction rates of nestin, a characteristic biological marker of neural stem cells, were higher at 2h and 10h after induction, which were 48% and 71.4% respectively, but only 0.06% at 48h after induction. Cells did not express GFAP after induction. The result of Nissl staining showed that Nissl body was contained in the cytoplasm after induction. The results of this study demonstrate that DMSO and BME can induce MSCs to obtain MSCs-derived neurons with neuronal phenotype.