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首页> 外文期刊>Einstein (So Paulo) >Evaluation of umbilical cord mesenchymal stem cell labeling with superparamagnetic iron oxide nanoparticles coated with dextran and complexed with Poly-L-lysine
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Evaluation of umbilical cord mesenchymal stem cell labeling with superparamagnetic iron oxide nanoparticles coated with dextran and complexed with Poly-L-lysine

机译：涂覆葡聚糖并与聚-L-赖氨酸复合的超顺磁性氧化铁纳米颗粒对脐带间充质干细胞标记的评价

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OBJECTIVE: The objective of this study was to evaluate the effect of the labeling of umbilical cord vein derived mesenchymal stem cells with superparamagnetic iron oxide nanoparticles coated with dextran and complexed to a non-viral transfector agent transfector poly-L-lysine. METHODS: The labeling of mesenchymal stem cells was performed using the superparamagnetic iron oxide nanoparticles/dextran complexed and not complexed to poly-L-lysine. Superparamagnetic iron oxide nanoparticles/dextran was incubated with poly-L-lysine in an ultrasonic sonicator at 37°C for 10 minutes for complex formation superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine by electrostatic interaction. Then, the mesenchymal stem cells were incubated overnight with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran. After the incubation period the mesenchymal stem cells were evaluated by internalization of the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran by Prussian Blue stain. Cellular viability of labeled mesenchymal stem cells was evaluated by cellular proliferation assay using 5,6-carboxy-fluorescein-succinimidyl ester method and apoptosis detection by Annexin V- Propidium Iodide assay. RESULTS: mesenchymal stem cells labeled with superparamagnetic iron oxide nanoparticles/dextran without poly-L-lysine not internalized efficiently the superparamagnetic iron oxide nanoparticles due to its low presence detected within cells. Mesenchymal stem cells labeled with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine efficiently internalized the superparamagnetic iron oxide nanoparticles due to greater presence in the cells interior. The viability and apoptosis assays demonstrated that the mesenchymal stem cells labeled and not labeled respectively with the superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine continue to proliferate over seven days and the percentage of cells in early or late apoptosis is low compared to the percentage of live cells over the three days. CONCLUSION: Our results showed that the use of poly-L-lysine complexed with superparamagnetic iron oxide nanoparticles/dextran provides better internalization of these superparamagnetic iron oxide nanoparticles in mesenchymal stem cells Thus, we demonstrated that this type of labeling is not cytotoxic to the mesenchymal stem cells, since the viability and apoptosis assays showed that the cells remain alive and proliferating. The efficiency of this type of labeling in mesenchymal stem cells can provide non-invasive methods for monitoring these cells in vivo.
机译：目的：本研究的目的是评估涂有右旋糖酐并与非病毒转染因子转染子聚L-赖氨酸复合的超顺磁性氧化铁纳米颗粒标记脐带静脉间充质干细胞的效果。方法：使用超顺磁性氧化铁纳米颗粒/葡聚糖复合或不复合聚-L-赖氨酸对间充质干细胞进行标记。将超顺磁性氧化铁纳米颗粒/右旋糖酐与聚-L-赖氨酸在超声仪中于37°C孵育10分钟，以通过静电相互作用复合形成超顺磁性氧化铁纳米颗粒/右旋糖酐/聚-L-赖氨酸。然后，将间充质干细胞与复杂的超顺磁性氧化铁纳米颗粒/葡聚糖/聚-L-赖氨酸和超顺磁性氧化铁纳米颗粒/葡聚糖孵育过夜。潜伏期后，通过普鲁士蓝染色内在的超顺磁性氧化铁纳米颗粒/葡聚糖/聚-L-赖氨酸和超顺磁性氧化铁纳米颗粒/葡聚糖的内在化来评估间充质干细胞。通过使用5，6-羧基-荧光素-琥珀酰亚胺酯法的细胞增殖测定和通过膜联蛋白V-碘化丙啶测定的凋亡检测，评价标记的间充质干细胞的细胞生存力。结果：用超顺磁性氧化铁纳米颗粒/右旋糖酐标记的不含聚-L-赖氨酸的间充质干细胞由于在细胞内的低存在而不能有效内化超顺磁性氧化铁纳米颗粒。由于细胞内部更多的存在，用复杂的超顺磁性氧化铁纳米颗粒/葡聚糖/聚-L-赖氨酸标记的间充质干细胞有效地内化了超顺磁性氧化铁纳米颗粒。生存力和凋亡测定表明，分别用超顺磁性氧化铁纳米颗粒/右旋糖酐/聚-L-赖氨酸标记和未标记的间充质干细胞持续增殖超过7天，与之相比，早期或晚期凋亡的细胞百分比较低三天内活细胞的百分比。结论：我们的研究结果表明，将聚-L-赖氨酸与超顺磁性氧化铁纳米颗粒/右旋糖酐配合使用，可以更好地将这些超顺磁性氧化铁纳米颗粒在间充质干细胞中内化。因此，我们证明了这种标记对间充质细胞没有细胞毒性干细胞，因为生存力和凋亡检测表明细胞保持存活和增殖。间充质干细胞中这种标记的效率可以提供非侵入性的方法来体内监测这些细胞。

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