不同诱导剂对大鼠骨髓间充质干细胞向神经细胞分化的作用

目的 观察体外不同诱导方法对大鼠骨髓间充质干细胞(MSCs)向神经细胞分化的作用.方法 体外培养、扩增骨髓MSCs,分别采用化学诱导剂(2-巯基乙醇 二甲基亚砜)和生物诱导剂,诱导骨髓MSCs分化为神经细胞.用神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF)和胶质纤维酸性蛋白(GFAP)免疫组织化学方法对诱导后的细胞进行鉴定和诱导率分析.应用组织学方法显示尼氏体.结果 倒置显微镜下可见,MSCs经两种方法诱导48h后,细胞均向神经细胞分化,胞体呈锥形、多角形,由胞体伸出较长的轴突样和树突样突起,且有分支.免疫组织化学鉴定显示,诱导后的细胞能特异性表达NSE、NF,不表达GFAP.化学诱导剂组NSE阳性细胞率为86.9%,NF阳性细胞率为85.6%;生物诱导剂组NSE阳性细胞率为88.2%,NF阳性细胞率为86.8%,两组间无显著差异(P＞0.05).硫堇-伊红染色显示胞体内有大量的尼氏体.结论 体外两种诱导方法均可诱导大鼠骨髓MSCs分化为神经细胞.

THE EFFECTS OF THE DIFFERENT INDUCERS ON THE DIFFERENTIATION OF RAT BONE-MARROW-DERIVED MESENCHYMAL STEM CELLS TO NEUROCYTES IN VITRO

Objective To observe the effects of the different inducers on the differentiation of bone-marrow-derived mesenchymal stem cells(MSCs) to neurocytes in vitro.Methods Isolation,cultivation and amplification of rat bone-marrow-derived MSCs were carried out,and the purified MSCs of the third generation were induced to differentiate into neurocytes by using chemical inducer and biological inducer,respectively.The expression of NSE,NF and GFAP of induced cells in two induced groups were showed using immunocytochemical method,and the induction rates in two groups were analyzed.The Nissl bodies in the induced cells were displayed by thionine-eosin staining.Results Under the convert microscope it was observed that the MSCs started to change their shape, and there were several axon-or dendrite-like processes out from the cell body when the MSCs were induced for 48hours.The induced cells derived from bone marrow MSCs expressed NSE and NF that are the special markers of the mature neurocytes,but they did not express GFAP.The reduction rates of NSE and NF in chemical inducer group were 86.9% and 85.6%,respectively,and in biological inducer group they were 88.2% and 86.8%, respectively.There were no significant differences between these two groups in reduction rates of NSE and NF(P>0.05).A lot of Nissl bodies in the cell body were seen in induced cells shown by thionine-eosin staining.Conclusion The bone-marrow-derived MSCs can differentiate into neurocytes by either chemical or biological inducers.