大鼠骨髓内皮祖细胞SPIO标记及检测

发布时间：2018-04-10 22:32

本文选题：内皮祖细胞 + 骨髓　；参考：《中国医药导报》2011年09期

【摘要】：目的:研究大鼠骨髓内皮祖细胞(endothelial progenitor cells,EPCS)的分离、培养、SPIO标记,SPIO对EPCS的标记效果及其对细胞存活、增殖以及凋亡的影响,为进一步的实验研究奠定基础。方法:SD大鼠体重120～150 g,采用梯度密度离心法和贴壁法获取大鼠EPCS,采用25μg/ml SPIO对其标记,比较标记和未标记的EPCS。普鲁士蓝染色观察铁标记率,台盼蓝检测细胞活力,MTT法检测细胞增殖力,Calcein-AM/PI染色检测细胞活性及细胞膜完整性,AO/PI染色检测细胞凋亡率。结果:分离培养EPCS 7～10 d,细胞集落相互连接,呈典型的铺路石样外观。普鲁士兰染色显示SPIO(25μg/ml,24 h)对细胞的标记率接近94%;比较SPIO标记及未标记的EPCS,细胞活力分别为(94.34±0.25)%和(95.33±0.31)%;MTT法检测两组间相比差异无统计意学义(P0.05);Calcein-AM/PI染色检测细胞的活性和膜完整性,存活率分别为96%和97%;AO/PI染色两组细胞的凋亡率均为5%。结论:采用梯度离心法从骨髓中分离单个核细胞,经诱导培养可实现内皮祖细胞的分离培养。25μg/ml浓度的SPIO细胞标记率高,对细胞毒性小,且不影响EPCS的活力、增殖及凋亡。
[Abstract]:Objective: to study the isolation of endothelial progenitor cells from rat bone marrow endothelial progenitor cells and the effect of Spio on EPCS labeling and its effects on cell survival, proliferation and apoptosis, so as to lay a foundation for further experimental study.Methods Sprague-Dawley (SD) rats weighing 120 ~ 150g were obtained by gradient density centrifugation and adherent method. 25 渭 g/ml SPIO was used to label the EPCSs, and the labeled and unlabeled EPCSs were compared.The rate of iron labeling was observed by Prussian blue staining, the cell viability was detected by trypan blue assay and the cell viability was detected by MTT assay. The cell activity was detected by Calcein-AM / Pi staining and the apoptosis rate was detected by AOP staining.Results: after EPCS was isolated and cultured for 7 ~ 10 days, the colony was connected with each other, showing a typical paving stone appearance.The survival rates were 96% and 97%, respectively, and the apoptotic rates were 5% in both groups.Conclusion: mononuclear cells were isolated from bone marrow by gradient centrifugation. The SPIO cells with 25 渭 g/ml concentration of endothelial progenitor cells could be isolated and cultured with high labeling rate, low cytotoxicity and no effect on the activity, proliferation and apoptosis of EPCS.
【作者单位】：山西医科大学第一医院影像科;
【基金】：山西医科大学青年科技研究基金(项目名称:SPIO标记干细胞移植治疗兔脊髓损伤的MR活体示踪项目编号:2010021038-2)
【分类号】：R329

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