促红细胞生成素基因修饰内皮祖细胞移植治疗下肢动脉闭塞：磁标记MR成像评价  

Transplantation of erythropoietin gene-modified endothelial progenitor cells to treat lower extremity artery occlusion: a magnetically-labeled MRI evaluation

作　　者：徐广宇[1] 田素红 周士琦 

机构地区：[1]唐山市协和医院CT/MRI室,河北省唐山市063000 [2]华北理工大学附属医院介入科,河北省唐山市063000

出　　处：《中国组织工程研究》2016年第41期6183-6189,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景：促红细胞生成素及内皮祖细胞移植均对下肢动脉闭塞有一定的治疗作用。目的：探讨超顺磁氧化铁纳米颗粒（super paramagnetic iron oxide,SPIO）标记的内皮祖细胞体外促红细胞生成素基因修饰效果及体外磁共振成像的可行性。方法：将对数生长期的大鼠骨髓来源内皮祖细胞分4组培养,内皮祖细胞组,SPIO标记转染组将pcD NA3-EPO重组质粒并转染至内皮祖细胞,随后进行SPIO标记;SPIO标记空载病毒组将空质粒转染至内皮祖细胞,随后进行SPIO标记;SPIO标记内皮祖细胞组直接进行SPIO标记。采用4.7 T MR成像SPIO标记的内皮祖细胞;检测4组细胞增殖、细胞周期分布及促红细胞生成素蛋白表达。结果与结论：（1）MR成像：T1WI、T2WI、T2＊WI序列均见细胞信号降低,随着细胞数目增多,信号降低逐渐明显;相同数量级细胞,T1WI信号降低最弱,T2＊WI信号降低最明显;T1WI、T2WI、T2＊WI所能检测到的最小细胞数分别为2×104、1×10^4、0.5×10^4;（2）细胞增殖、细胞周期分布：3组标记内皮祖细胞增殖、细胞周期分布与内皮祖细胞组比较差异均无显著性意义;（3）促红细胞生成素蛋白表达：仅SPIO标记转染组可见促红细胞生成素蛋白表达;（4）结果表明：SPIO标记的内皮祖细胞体外促红细胞生成素基因修饰后对细胞增殖、细胞周期无影响,4.7 T MR能够在体外对SPIO标记的促红细胞生成素基因修饰内皮祖细胞成像。BACKGROUND： Erythropoietin and progenitor cell transplantation both have therapeutic effects on lower extremity arterial occlusive disease.OBJECTIVE： To investigate the erythropoietin modification effect and magnetic resonance imaging feasibility of superparamagnetic iron oxide（SPIO）-labeled endothelial progenitor cells in vitro. METHODS： Rat bone marrow-derived endothelial progenitor cells at logarithmic growth phase were randomized into four groups： endothelial progenitor cell group, SPIO labeled transfection group（pc DNA3-EPO transfection followed by SPIO labeling）, SPIO labeled empty vector group（empty plasmid transfection followed by SPIO labeling）, and SPIO labeling group（only SPIO labeling）. 4.7T MRI was used to observe SPIO-labeled endothelial progenitor cells. Cell proliferation, cell cycle distribution, and expression of erythropoietin protein in the four groups were measured. RESULTS AND CONCLUSION： MRI findings showed with the increasing cell number, gradually lowered signal intensity on T1-weighted imaging（T1WI）, T2 WI and T2＊WI was seen, and the reduction in the signal intensity was the maximum on T2＊WI sequence and the minimum on T1 WI sequence. For T1 WI, T2 WI and T2＊WI sequences, the minimum number of cells was 2×10^4, 1×10^4 and 0.5×10^4, respectively. Cell proliferation and cell cycle distribution showed no significant difference among three SPIO labeling groups. In addition, the expression of erythropoietin protein was only found in the SPIO-labeled transfection group. These findings showed that under SPIO labeling, erythropoietin gene-modified endothelial progenitor cells show no changes in cell proliferation and cell cycle, and the 4.7T MR is capable of imaging SPIO-labeled erythropoietin gene-modified endothelial progenitor cells in vitro.

关 键 词：干细胞 移植 内皮祖细胞 超顺磁性氧化铁粒子 促红细胞生成素 基因修饰 MR 下肢动脉闭塞 

分 类 号：R394.2[医药卫生—医学遗传学]