Feasibility of lentiviral-mediated sodium iodide symporter gene delivery for the efficient monitoring of bone marrow-derived mesenchymal stem cell transplantation and survival
Feasibility of lentiviral-mediated sodium iodide symporter gene delivery for the efficient monitoring of bone marrow-derived mesenchymal stem cell transplantation and survival
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DOI：
10.3892/ijmm.2014.1970
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发表时间：
2014-12-01
影响因子：
5.4
通讯作者：
Li, Biao
中科院分区：
文献类型：
--
作者：
Shi, Shuo;Zhang, Min;Li, Biao
本研究旨在探讨慢病毒介导的钠碘转运体（NIS）基因转染监测骨髓间充质干细胞（BMSC）移植入梗死心肌的可行性。为此，我们构建了表达NIS和增强型绿色荧光蛋白（EGFP）的慢病毒载体（Lv-EF 1 alpha-NIS-IRES-EGFP），并将其以不同感染复数（MOI）导入BMSCs。荧光显微镜下观察EGFP的表达。在体外动态监测Lv-EF 1 α-NIS-IRES-EGFP处理的BMSC中的碘摄取和高氯酸钠（NaClO 4）对碘摄取的抑制。将Lv-EF 1 α-NIS-IRES-EGFP处理的BMSCs移植到Sprague-Dawley大鼠的梗死心肌中，并在移植后1周在体内进行Tc-99 m（99 g）（Tc，锝; 99 m表示锝处于其激发阶段; 99 g表示锝的原子量）微型单光子发射计算机断层扫描（SPECT）/计算机断层扫描（CT）成像。分离的骨髓间充质干细胞成功分化为脂肪细胞和成骨细胞。骨髓基质细胞表面标志CD 105、CD 29和CD 90阳性，CD 14、CD 34和CD 45阴性。Lv-EF 1 α-NIS-IRES-EGFP可有效转染BMSCs。RT-qPCR和western blot分析证实，BMSCs在感染后第7天表达高水平的NIS蛋白和mRNA，从第14天到第21天NIS表达保持在一致的水平。在Lv-EF 1 α-NIS-IRES-EGFP处理的BMSC中，在体外观察到碘-125（I-125）的积累，并且在移植后1周通过Tc-99 m（99 g）micro-SPECT/CT成像成功地监测。这些结果表明，慢病毒载体是研究BMSCs基因传递的有力工具。利用慢病毒载体介导NIS基因用于无创性监测BMSC移植和在梗死心肌中存活是可行的。
The aim of the present study was to explore the feasibility of lentiviral-mediated sodium iodide symporter (NIS) gene delivery for monitoring bone marrow-derived mesenchymal stem cell (BMSC) transplantation into the infarcted myocardium. For this purpose, we constructed a lentiviral vector (Lv-EF1 alpha-NIS-IRES-EGFP) expressing NIS and enhanced green fluorescent protein (EGFP), and introduced it into BMSCs at different multiplicities of infection (MOI). The expression of EGFP was observed under a fluorescence microscope. Iodine uptake and the inhibition of iodine uptake by sodium perchlorate (NaClO4) in the Lv-EF1 alpha-NIS-IRES-EGFP-treated BMSCs were dynamically monitored in vitro. The Lv-EF1 alpha-NIS-IRES-EGFP-treated BMSCs were transplanted into the infarcted myocardium of Sprague-Dawley rats, and Tc-99m(99g) (Tc, technetium; 99m indicates that technetium is at its excited stage; 99g indicates the atomic weight of technetium) micro-single-photon emission computed tomography (SPECT)/computed tomography (CT) imaging was performed in vivo 1 week following transplantation. The isolated BMSCs successfully differentiated into adipocytes and osteoblasts. The BMSCs were positive for the cell surface markers, CD105, CD29 and CD90, and negative for CD14, CD34 and CD45. Lv-EF1 alpha-NIS-IRES-EGFP was efficiently transfected into the BMSCs. RT-qPCR and western blot analysis confirmed that the BMSCs expressed high protein and mRNA levels of NIS by day 7 following infection, and NIS expression remained at a consistent level from day 14 to 21. In the Lv-EF1 alpha-NIS-IRES-EGFP-treated BMSCs, the accumulation of iodine-125 (I-125) was observed in vitro and was successfully monitored by Tc-99m(99g) micro-SPECT/CT imaging at 1 week following transplantation. These results suggest that lentiviral vectors are powerful vehicles for studying gene delivery in BMSCs. It is feasible to use lentiviral vectors to deliver an NIS gene for the non-invasive monitoring of BMSC transplantation and survival in the infarcted myocardium in vivo.