BHA诱导人脐带间充质干细胞分化为神经细胞-东南大学学报医学版

BHA诱导人脐带间充质干细胞分化为神经细胞

单位：1. 湖北十堰市太和医院核医学科,湖北十堰 442000;
2. 湖北十堰市太和医院生命科学研究所,湖北十堰 442000

分类号：R329.2

出版年·卷·期（页码）：2013·32·第二期（201-205）

摘要：

目的:探讨丁基羟基茴香醚(BHA)能否将人脐带间充质干细胞(hUC-MSCs)诱导分化为神经细胞。方法:正常培养的第4代hUC-MSCs利用200 μmol·L^-1 BHA加2 mmol·L^-1巯基乙醇(β-ME)进行诱导。每12 h观察细胞形态变化,并于诱导后24和72 h收集细胞,提取RNA,RT-PCR检测基因表达情况,于第3天进行免疫荧光染色。结果:诱导12 h后即有少量细胞发生形态变化,随着时间的推移变化越来越明显,72 h时绝大部分细胞都已变成神经细胞形态。免疫荧光染色nestin、GFAP及NSE阳性。RT-PCR检测诱导后24 h开始有nestin、MAP及NF-L表达,72 h后nestin和NF-L表达减少,MAP表达增强,同时NeuroD1也有较强表达,表明此时已分化为成熟的神经元细胞。结论:BHA能快速地将hUC-MSCs诱导分化为神经细胞。

Objective: To investigate whether butyl hydroxyanisole (BHA) and β-mereaptoethanol(β-ME) could be used to induce human umbilical cord mesenchymal stem cells(hUC-MSCs) into neural cells.Methods: The normal cultured fourth passage hUC-MSCs were randomly divided into experiment group and control group.The experiment group were induced by 200 μmol·L^-1 BHA+2 mmol·L^-1 β-ME, and the control group were continuously cultured with common MSC medium.The morphologic changes of cells were observed every 12 hours. The cells of the two groups were collected after 24 h and 72 h distinctively to extract total RNA and analyzed neural cell specific genes by RT-PCR.On the third day after induction, the expressions of nestin,glial fibrillary acidic protein(GFAP) and neuron-specific enolase(NSE) were detected by immunohistochemistry.Results: Within 12 hours of induction,some of the hUC-MSCs assumed neuronal morphology,the cell became smaller, diacaustic was higher.These changes became more and more obvious as the time being.After induction for 72 h,a large number of responsive cells exhibited typical neuronal morphology.Nestin,GFAP and NSE were immunepositive.RT-PCR indicated that nestin,MAP and NF-L began to express 24 h after induction.72 h later,the expression of nestin and NF-L were reduced,while MAP and NeuroD1 expression became increased.Conclusion: BHA can induce hUC-MSCs differentiated into neural cells quickly.

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