隐丹参酮诱导骨髓间质干细胞分化为神经元样细胞的实验研究

作者	单位
夏文杰	中山医科大学病理生理教研室
张秀明	中山医科大学附属第一医院心胸外科
李艳	广东药学院病理生理教研室
李树浓	美国芝加哥大学人类遗传研究所
陈振光	中山医科大学病理生理教研室
张丽蓉	中山医科大学病理生理教研室
项鹏	中山医科大学病理生理教研室

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中文摘要:

目的:观察隐丹参酮体外定向诱导成人骨髓间质干细胞(MSC)分化为神经元样细胞。方法:采用Ficoll-Paque液分离成人MSC,体外扩增,FACScan流式细胞仪检测表面抗原表达,采用含隐丹参酮的无血清DMEM诱导MSC分化为神经元样细胞。免疫组化鉴定神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF—M,NF—H)、胶质纤维酸性蛋白(GFAP)、巢蛋白(Nestin)的表达。结果:成人骨髓间质干细胞在体外扩增原代可获得0.5×106个细胞,15代可获得(2～3)×1012个细胞,流式细胞仪检测显示CD29、CD44、CD90、CD105、CD166表达阳性,CD11a、CD14、CD34、CD38、CD45、CD80、CD86为阴性。加入隐丹参酮诱导后,MSC胞体收缩,突起伸出;免疫组化显示诱导出的神经元样细胞NSE、NF-M,NF-H、Nestin表达阳性.GFAP阴性。结论:隐丹参酮在体外可诱导成人骨髓间质干细胞分化为神经元样细胞。

英文摘要:

To investigate the effect of cryptotanshjnone (CT) in inducing oriental differentiation of human rib bone marrow mesenchymal stem cells (MSC) to neuron-like cells. Methods: Adult MSC were separated with Ficoll-Paque reagent and amplified in culture medium. The surface antigen was detected by FACScan flow cy-tometry. MSCs were induced to differentiate into neurons with Dulbecco’s modified Eagle’s medium (DMEM) containing CT, and neuron specific enolase (NSE), neurofilament (NF-M, NF-H), nestin, glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. Results: By amplifying in vitro, 0.5 x 106 human adult bone marrow MSC of primary passage and 2 - 3 x 1012 of 15 passages could be obtained. Flow cyto-metric examination showed positive expression of CD29, CD44, CD90, CD105, CD166 and negative expression of GDI la, CD14, CD34, CD38, CD45, CD80, and CD86. After induced by CT, the cytoplasm of MSC contracted with protruding process. Immunohistochemical examination displayed the neuron-like cells with the NF-M, NF-H and nestin expression positive, and negative expression of GFAP. Conclusion: CT could induce human adult bone marrow MSC differentiated to neuron-like cells.

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