Fth1基因标记对骨髓间充质干细胞生物学特性及体外MRI表现的影响  

Effect of Fth1gene on biological characteristics and MRI feature of bone marrow mesenchymal stem cells in vitro

作　　者：薛杨[1] 黄晓蕾[1] 蔡伶伶[1] 王博成[1] 詹青[2] 赵江民[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院影像科,上海201999 [2]上海中医药大学附属第七人民医院影像科,上海200137

出　　处：《第二军医大学学报》2016年第7期841-845,共5页Academic Journal of Second Military Medical University

基　　金：上海市科委科技创新行动计划重点项目(10411953400);上海交通大学医学院项目(12XJ30061);上海市卫生局科研课题(20124194);上海市宝山区科学技术委员会科研项目(14-E-4)~~

摘　　要：目的观察铁蛋白报告基因Fth1标记对骨髓间充质干细胞(MSCs)生物学特性及体外MRI表现的影响。方法原代分离培养大鼠MSCs,取第4代MSCs,用构建的携带铁蛋白重链基因Fth1的慢病毒载体进行感染,并在培养液内加入1mol/L柠檬酸铁进行培养。采用普鲁士蓝染色检测MSCs的摄铁能力,锥虫蓝染色活细胞计数检测MSCs的细胞存活率,CCK-8方法测定MSCs的增殖活性。应用MRI的FSE T2WI和SWAN序列观察Fth1标记的MSCs和未标记MSCs MRI信号的差异。结果 Fth1基因可成功转染MSCs,转染MSCs的普鲁士蓝染色标记效率为87%。未加含铁培养液的Fth1标记MSCs细胞存活率为(92.17±1.91)%,细胞增殖活性D值为1.094±0.068,与未标记MSCs比较差异均无统计学意义[细胞存活率为(94.23±2.42)%,细胞增殖活性D值为1.027±0.122;P>0.05);加入含铁培养液培养3d后的Fth1标记MSCs细胞存活率为(77.47±4.10)%,细胞增殖活性D值为0.493±0.024,与未加含铁培养液的Fth1标记MSCs及未标记MSCs比较差异均有统计学意义(P<0.05)。MRI扫描FSE T2WI和SWAN序列显示,加入含铁培养液培养5d后Fth1标记MSCs信号强度为656.6±18.2,与未加含铁培养液的Fth1标记MSCs及未标记MSCs比较差异均有统计学意义(分别为807.3±17.1和847.1±10.5,P<0.05),而未加含铁培养液的Fth1标记MSCs与未标记MSCs比较差异无统计学意义(P>0.05)。结论 Fth1报告基因可成功转染MSCs,转染后的MSCs可高效表达并摄取铁;细胞活力及增殖活性在不含铁的培养液中不受影响,但在铁浓度1mol/L含铁培养液中受到一定影响;经含铁培养液培养5d后,MRI可体外检测Fth1标记MSCs,其FSE T2WI和SWAN序列呈低信号改变。Objective To observe the effect ferritin reporter gene Fth1 labeling on biological characteristics and magnetic resonance imaging（MRI）findings of bone marrow mesenchymal stem cells（MSCs）in vitro.Methods Rat MSCs were isolated and cultured by plastic adherence method,and the lentiviral vectors carrying the ferritin heavy chain gene Fth1 were constructed to transfect the 4th passage MSCs transfected with Fth1,and the MSCs were cultured in the medium added with 1mol/L ferric citrate.Prussian blue staining was used to evaluate the iron uptake ability of the transfected MSCs,trypan blue staining was used to evaluate cell viability,and CCK-8assay was used to evaluate the proliferation activity of transfected MSCs.The MRIsignal differences between transfected MSCs and normal MSCs were observed by MRI FSE T2 WI and SWAN sequence.Results MSCs were successfully transfected with Fth1 gene,with the Prussian blue staining efficiency being 87%.For transfected MSCs without ferric citrate medium,the cell survival rate was（92.17±1.91）% and absorbance（D）value was1.094±0.068,which were not significantly different from those of the normal MSCs（[94.23±2.42]% and 1.027±0.122,P〉0.05）.For transfected MSCs treated with ferric citrate medium for 3days,the cell survival rate was（77.47±4.10）% and Dvalue was 0.493±0.024,which were significantly different from those without ferric citrate treatment（P〈0.05）.The signal strength of MRI FSE T2 WI and SWAN sequence for transfected MSCs treated with ferric citrate medium was 656.6±18.2,which was significantly different from without ferric citrate treatment and the normal MSCs（807.3±17.1and 847.1±10.5,P〈0.05）;and difference between without ferric citrate treatment and the normal MSCs was not statistically significant（P〉0.05）.Conclusion MSCs can be successfully transfected with Fth1 reporter gene,and the transfected MSCs can efficiently overexpress and uptake iron.The cell viability and proliferation is not affected in ferric citrate free medium,bu

关 键 词：铁蛋白报告基因 骨髓间充质干细胞 生物学特性 磁共振成像 

分 类 号：R445.2[医药卫生—影像医学与核医学]