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用超顺磁性氧化铁纳米颗粒对从外周血中分离的内皮祖细胞进行体外标记。

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

机构信息

Key Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health, Department of Surgery, First Affiliated Hospital, School of Medicine, Zhejiang, PR China.

出版信息

Mol Med Rep. 2012 Aug;6(2):282-6. doi: 10.3892/mmr.2012.912. Epub 2012 May 10.

Abstract

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.

摘要

内皮祖细胞(EPCs)的移植为治疗人类肿瘤或血管疾病提供了一种新方法。磁共振成像(MRI)已被证明通过超顺磁氧化铁(SPIO)纳米粒子标记细胞在追踪移植的干细胞方面非常有效。SPIO 已被用于标记和追踪 EPCs;然而,SPIO 对 EPCs 的细胞水平影响尚不清楚。在本研究中,我们在体外使用自制的 SPIO 纳米粒子标记 EPCs,并评估标记 EPCs 的生物学特性。从新西兰兔外周血中分离 EPCs,并在纤连蛋白包被的培养瓶中培养。将 EPCs 用终铁浓度为 20μg/ml 的自制 SPIO 纳米粒子标记。用透射电子显微镜和普鲁士蓝染色证实标记的 EPCs。通过原子吸收光谱法检测铁/细胞量。通过流式细胞术分析检测 EPCs 的膜抗原。比较标记和未标记的 EPCs 之间的细胞活力和增殖能力。兔 EPCs 被有效标记,标记效率约为 95%。SPIO 纳米粒子定位于 EPCs 的内体小泡中,这通过透射电子显微镜得到证实。标记和未标记的 EPCs 之间的细胞活力和增殖能力没有显著差异(P>0.05)。结论:自制 SPIO 纳米粒子有效标记了兔外周血 EPCs,而不影响其主要生物学特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90d5/3493051/58f11390c6f1/MMR-06-02-0282-g00.jpg

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