骨髓间充质干细胞诱导分化为神经细胞的表型变化

Phenotype features of neurocyte from bone marrow-derived mesenchymal stem cells

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。

In order to investigate the phenotype features of neurocyte from bone marrow-derived mesenchymal stem cells （ MSCs）, the rat MSCs from bone marrow were cultured by adhering to the culture plasticdish. The cultures were induced by the chemical inducers of the combination of dimethyl sulfoxide （DMSO） and β-Mercaptoethanol （BME）. Then the expressions of nestin, which is the special marker of neural stem cells, MAP2 ,NSE and NF, which are the markers of mature neurocyte of induced cells, and GFAP, which is the marker of the glial cells, were detected using immunocytochemical method. The Nissl bodies in the induced ceils were showed by thionine-eosin staining. The results showed that after the MSCs were induced into neurencytes by DMSO ＋ BME for 48 h, 80% of the induced MSCs trans- formed into neuron-like cells. There were several processes ecphyma from the cell body, and their processes liked axons or dendrites. The induced cells expressed MAP2, NSE and NF and the expression rates of them are 88.6%, 87.1% and 85.8%, respectively. The nestin expression rates of induced MSCs in 2 h and 10 h were 48% and 71.4%, respectively, which were higher than that of induced MSCs in 48 h （0.06%）. The induced MSCs did not express GFAP. Lots of Nissl bodies were presented in the cytoplasms of the induced cells. The present results suggest that the neurocytes which express phenotype features of neuron can be obtained from bone marrow MSCs by induction of DMSO ＋ BME.