Baculovirus as an ideal radionuclide reporter gene vector: a new strategy for monitoring the fate of human stem cells in vivo.
Baculovirus as an ideal radionuclide reporter gene vector: a new strategy for monitoring the fate of human stem cells in vivo.
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DOI：
10.1371/journal.pone.0061305
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Pan Y;Liu S;Wu H;Lv J;Xu X;Zhang Y
放射性核素报告基因成像有望用于移植干细胞的无创监测。因此，利用重组杆状病毒携带钠碘转运体（NIS）报告基因的放射性核素显像监测干细胞治疗的可行性进行了探讨。构建了携带NIS和绿色荧光蛋白（GFP）报告基因的重组杆状病毒（Bac-NIS和Bac-GFP），分别感染人诱导多能干细胞（hiPSCs）、人胚胎干细胞（hESC）和人脐带血间充质干细胞（hUCB-MSCs）。通过流式细胞术测定感染效率、总荧光强度和转基因表达的持续时间。使用CCK-8试验评估了杆状病毒对hUCB-MSC的细胞毒性/增殖作用。在Bac-NIS感染的hUCB-MSC上进行125 I摄取和高氯酸盐抑制测定。通过NanoSPECT/CT成像对移植有Bac-NIS感染的hUCB-MSC的小鼠进行放射性核素成像。重组杆状病毒在hESC、hiPSC和hUCB-MSC中的感染效率随着MOI的增加而增加（在MOI = 800时，分别为27.3%、35.8%和95.6%）。 几乎没有观察到细胞毒性，仅对hUCB-MSCs增殖有轻微影响。感染后8天仍有明显的GFP表达（40.6%）。放射性碘的功能积累的NIS基因产物和高氯酸盐（ClO 4-）的特异性抑制。放射性碘摄取，在30分钟达到峰值，并随着时间的推移逐渐下降，与hUCB-MSC细胞数量显着相关（R2 = 0.994）。 最后，放射性核素成像显示Bac-NIS感染的hUCB-MSCs在体内有效地积累放射性碘，随着时间的推移逐渐减弱。杆状病毒作为转基因载体的放射性核素报告基因显像技术是一种很有前途的干细胞移植治疗监测策略。
Radionuclide reporter gene imaging holds promise for non-invasive monitoring of transplanted stem cells. Thus, the feasibility of utilizing recombinant baculoviruses carrying the sodium iodide symporter (NIS) reporter gene in monitoring stem cell therapy by radionuclide imaging was explored in this study. Recombinant baculoviruses carrying NIS and green fluorescent protein (GFP) reporter genes (Bac-NIS and Bac-GFP) were constructed and used to infect human induced pluripotent stem cells (hiPSCs), human embryonic stem cells (hESCs) and human umbilical cord blood mesenchymal stem cells (hUCB-MSCs). Infection efficiency, total fluorescence intensity and duration of transgene expression were determined by flow cytometry. Cytotoxicity/proliferative effects of baculovirus on hUCB-MSCs were assessed using CCK-8 assays. 125I uptake and perchlorate inhibition assays were performed on Bac-NIS-infected hUCB-MSCs. Radionuclide imaging of mice transplanted with Bac-NIS-infected hUCB-MSCs was performed by NanoSPECT/CT imaging. Infection efficiencies of recombinant baculovirus in hESCs, hiPSCs and hUCB-MSCs increased with increasing MOIs (27.3%, 35.8% and 95.6%, respectively, at MOI = 800). Almost no cytotoxicity and only slight effects on hUCB-MSCs proliferation were observed. Obvious GFP expression (40.6%) remained at 8 days post-infection. The radioiodide was functionally accumulated by NIS gene products and specifically inhibited by perchlorate (ClO4 -). Radioiodide uptake, peaking at 30 min and gradually decreasing over time, significantly correlated with hUCB-MSCs cell number (R2 = 0.994). Finally, radionuclide imaging showed Bac-NIS-infected hUCB-MSCs effectively accumulated radioiodide in vivo, which gradually weakened over time. Baculovirus as transgenic vector of radionuclide reporter gene imaging technology is a promising strategy for monitoring stem cell transplantation therapy.
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影响因子：
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作者：
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发表时间：
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影响因子：
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