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In vitro labeling of endothelial progenitor cells isolated from peripheral blood with superparamagnetic iron oxide nanoparticles

机译:超顺磁性氧化铁纳米粒子体外标记从外周血中分离的内皮祖细胞

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摘要

The transplantation of endothelial progenitor cells (EPCs) provides a novel method for the treatment of human tumors or vascular diseases. Magnetic resonance imaging (MRI) has proven to be effective in tracking transplanted stem cells by labeling the cells with superparamagnetic iron oxide (SPIO) nanoparticles. The SPIO has been used to label and track the EPCs; however, the effect of SPIO upon EPCs remains unclear on a cellular level. In the present study, EPCs were labeled with home-synthesized SPIO nanoparticles in vitro and the biological characteristics of the labeled EPCs were evaluated. The EPCs were isolated from the peripheral blood of New Zealand rabbits and cultured in fibronectin-coated culture flasks. The EPCs were labeled with home-synthesized SPIO nanoparticles at a final iron concentration of 20 μg/ml. Labeled EPCs were confirmed with transmission electron microscopy and Prussian blue staining. The quantity of iron/cell was detected by atomic absorption spectrometry. The membranous antigens of EPCs were detected by cytofluorimetric analysis. Cell viability and proliferative capability between the labeled and unlabeled EPCs were compared. The rabbit EPCs were effectively labeled and the labeling efficiency was approximately 95%. The SPIO nanoparticles were localized in the endosomal vesicles of the EPCs, which were confirmed by transmission electron microscopy. No significant differences were found in cell viability and proliferative capability between labeled and unlabeled EPCs (P>0.05). In conclusion, rabbit peripheral blood EPCs were effectively labeled by home-synthesized SPIO nanoparticles, without influencing their main biological characteristics.
机译:内皮祖细胞(EPC)的移植提供了一种治疗人类肿瘤或血管疾病的新方法。事实证明,磁共振成像(MRI)通过用超顺磁性氧化铁(SPIO)纳米粒子标记细胞,可以有效地追踪移植的干细胞。 SPIO已用于标记和跟踪EPC;但是,SPIO对EPC的影响在细胞水平上尚不清楚。在本研究中,体外用家庭合成的SPIO纳米颗粒标记EPC,并评估标记的EPC的生物学特性。从新西兰兔的外周血中分离出EPC,并在纤连蛋白包被的培养瓶中培养。用最终合成铁浓度为20μg/ ml的家庭合成SPIO纳米颗粒标记EPC。通过透射电子显微镜和普鲁士蓝染色确认标记的EPC。通过原子吸收光谱法检测铁/电池的量。通过细胞荧光分析检测EPC的膜抗原。比较了标记的和未标记的EPC之间的细胞活力和增殖能力。兔EPC被有效标记,标记效率约为95%。 SPIO纳米颗粒位于EPC的内体囊泡中,这已通过透射电子显微镜确认。标记的和未标记的EPCs在细胞活力和增殖能力上均未发现显着差异(P> 0.05)。总之,家兔合成的SPIO纳米颗粒可有效标记兔外周血EPC,而不会影响其主要生物学特性。

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