骨髓间充质干细胞成神经分化中的FTH1基因表达

发布时间：2018-04-06 23:29

本文选题：间充质干细胞　切入点：神经诱导分化　出处：《第三军医大学学报》2016年19期

【摘要】：目的明确骨髓间充质干细胞(mesenchymal stem cells,MSCs)成神经分化对细胞内铁蛋白重链(ferritin heavy chain 1,FTH1)基因表达的影响,探讨FTH1报告基因成像检测定向分化细胞的可行性。方法以慢病毒为载体将FTH1基因转入MSCs中,利用全反式维甲酸和神经细胞诱导液对其诱导分化,Western blot检测分化前后细胞内FTH1的表达,MRI观察细胞T2WI信号变化,普鲁士蓝染色和透射电镜检测细胞内FTH1表达所引起的聚铁效应。结果携带FTH1基因的MSCs(MSCs-FTH1)成功分化为神经元样细胞(Neurons-FTH1)。Western blot检测显示MSCs-FTH1及Neurons-FTH1细胞中FTH1均明显表达;在500μmol/L枸橼酸铁铵的培养条件下,两种细胞MRI检查均发现T2WI信号明显降低;普鲁士蓝染色和透射电镜证实细胞内较多铁颗粒聚集。结论 MSCs成神经分化对细胞内FTH1基因的表达无明显影响,基于FTH1的MRI报告基因成像可用于MSCs神经分化后的检测。
[Abstract]:Objective to investigate the effect of mesenchymal stem cells (MSCs) neural differentiation on the expression of ferritin heavy chain 1 (FTH1) gene in bone marrow mesenchymal stem cells (BMSCs), and to explore the feasibility of FTH1 reporter gene imaging for the detection of differentiated cells.Methods the FTH1 gene was transfected into MSCs with lentivirus vector. The expression of FTH1 in the cells before and after differentiation was detected by using all-trans retinoic acid (ATRA) and neuronal inducer to detect the expression of FTH1 in the cells before and after differentiation. The changes of T2WI signal were observed.Prussian blue staining and transmission electron microscopy (TEM) were used to detect the polyferric effect induced by FTH1 expression.Results the expression of FTH1 in MSCs-FTH1 and Neurons-FTH1 cells was detected by Western blot. The results showed that the expression of FTH1 in both MSCs-FTH1 and Neurons-FTH1 cells was significantly lower than that in cultured cells with 500 渭 mol/L ammonium citrate, and the T2WI signal was significantly decreased by MRI examination of the two cells under the condition of culture of 500 渭 mol/L ammonium citrate, and the results of MRI analysis showed that the expression of FTH1 in both cells was significantly lower than that in the control group (P < 0.05).Prussian blue staining and transmission electron microscopy (TEM) showed that more iron particles were accumulated in the cells.Conclusion MSCs neural differentiation has no significant effect on the expression of FTH1 gene in cells. MRI reporter gene imaging based on FTH1 can be used for the detection of MSCs neural differentiation.
【作者单位】：重庆医科大学附属儿童医院放射科;儿童发育疾病研究省部共建教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地;
【基金】：国家自然科学基金面上项目(81171387)~~
【分类号】：R329.2

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