丹参素定向诱导骨髓间质干细胞分化为神经元样细胞的研究

作者	单位
余勤	浙江中医学院
罗依	浙江大学附属第一医院
鄂艳	浙江中医学院
盛丽先	浙江中医学院
董勤	浙江省中医院
丁伟	浙江大学附属第一医院
郭莹	浙江中医学院

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中文摘要:

目的研究丹参素体外定向诱导骨髓间质干细胞 (mesenchymalstemcells,MSC)分化为神经元样细胞。方法利用Ficoll Paque液将骨髓细胞进行密度梯度离心和贴壁筛选分离出MSC ,体外扩增 ,流式细胞仪检测MSC表面抗原表达。采用含丹参素的无血清L DMEM诱导MSC分化为神经元样细胞 ,并且探讨丹参素不同浓度及不同作用时间对MSC诱导分化为神经元样细胞的影响。以免疫组化方法检测神经元特异性烯醇化酶 (NSE)、神经丝蛋白 (NF M )、巢蛋白 (nestin)、胶质纤维酸性蛋白 (GFAP)的表达。结果MSC在体外传代扩增后 ,流式细胞仪检测结果显示CD2 9、CD4 4、CD16 6表达阳性 ,CD14、CD34、CD4 5、HLA DR为阴性。丹参素诱导后 ,MSC胞体收缩 ,突起伸出 ,呈典型的核周体形态 ,类似神经元 ;免疫组化显示诱导出的神经元样细胞NSE、NF M、nestin表达阳性 ,GFAP阴性。结论丹参素可以在体外诱导MSC分化为神经元样细胞 ,其诱导转化率与丹参素浓度和作用时间有关。

英文摘要:

Objective To study the effect of Danshensu (DSS) on directional differentiation of mesenchymal stem cells (MSC) into neuron like cells. Methods MSC were separated from bone marrow with density gradient centrifugation, wall sticking screening and amplified in vitro. Flow cytometry was used to monitor the expression of surface antigens. DSS contained in non serum L DMEM was used to induce differentiation of MSC to neuron like cells, and the effect of DSS when different concentration and acting time used was explored. And levels of neuron specific enolase (NSE), neurofilament protein (NF M), nestin, and expression of glial fibrillary acidic protein (GFAP) were measured by immunohistochemical method. Results After being propagated and amplified in vitro, MSC were positively expressed for CD29, CD44, CD166, and negatively expressed for CD14, CD34, CD45、HLA DR. After induction of DSS, MSC exhibited the typical form of perikaryon with pyknotic cell body and prominence projected like that of neuron. These cells were positively expressed in NSE, NF M and nestin, and negatively expressed in GFAP. Conclusion DSS could induce differentiation of MSC to neuron like cells in vitro, the action is concentration and time dependent.

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