骨髓间充质干细胞成神经分化中的FTH1基因表达  

Expression of ferritin heavy chain 1 in neural differentiation of bone marrow mesenchymal stem cells

作　　者：钟毅[1,2] 蔡金华[1] 贺小娅 秦勇[1] 廖一凡[1,2] 

机构地区：[1]重庆医科大学附属儿童医院放射科,重庆400014 [2]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

出　　处：《第三军医大学学报》2016年第19期2121-2127,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81171387)

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。Objective To determine the effect of neural differentiation of bone marrow mesenchymal stem cells （MSCs） on the expression of the reporter gene, ferritin heavy chain 1 （FTH1）, and to explore the feasibility of visualizing the neural differentiated cells with FTH1 for magnetic resonance reporter gene imaging. Methods The reporter gene FTH1 was transfected into the MSCs by using a lentivirus system. Then the MSCs carrying FTH1 （ MSCs-FTH1 ） were induced into a neural-direction differentiation with all- trans retinoic acid and modified neural induction media. The dynamic expression of the FTH1 in MSCs-FTH1 and neurons-FTH1 （the neural differentiated ceils ） were assessed by Western blotting. MR imaging was performed to detect the changes of T2WI signal intensity of both cells. Prussian blue staining and transmission electron microscopy were used to determine the iron accumulation in the cells. Results The MSCs-FTH1 successfully differentiated into neuron-like cells （Neurons-FTH1）. The expression of FTH1 was detected in both MSCs-FTH1 and Neurons-FFH1 by Western blotting. Under the cultural condition of 500 μmol/L of ferric ammonium citrate, remarkable decrease of T2WI signal was seen in the both cells. Prussian blue staining and transmission electron microscopy verified the extensive iron accumulation in MSCs-FTH1 and Neurons- FTH1. Conclusion The neural differentiation of MSCs has no impact on the expression of FTH1, therefore MRI based on the reporter gene FTH1 can be used in the detection of neural differentiated cells.

关 键 词：间充质干细胞 神经诱导分化 FTH1 磁共振成像 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]