中国医学科学院学报2012，Vol.34Issue(5)：474-479,6.DOI:10.3881/j.issn.1000-503X.2012.05.007
双模态标记F344大鼠骨髓间充质干细胞移植入大鼠心脏后离体及在体影像示踪
In Vivo and In Vitro Imaging Tracing of Dual-labeled Bone Mesenchymal Stem Cells Transplanted into Myocardium of F344 Rats
摘要
Abstract
Objective To investigate the feasibility of in vitro and in vivo magnetic resonance imaging ( MRI) and fluorescence imaging tracking of transplanted bone mesenchymal stem cells ( BMSCs) dual-labeled with ultrasmall superparamagnetic iron oxide (USPIO) and red fluorescence protein (RFP). Methods BMSCs were incubated with culture medium containing USPIO for 24 hours. The Prussian-blue staining, transmission electron microscopy and trypan-blue staining were used to study the efficacy and safety of labeling. F344 rat model of acute myocardial infarction was established by ligating the left anterior descending coronary artery. The dual-labeled BMSCs were injected into the margin of the infraction myocardium. Then MRI and fluorescence imaging were performed to trace the cells both in vitro and in vivo. Postmortal study was carried out to observe the distribution of transplanted cells in myocardium. Results The percentage of dual-labeled BMSCs reached 99% after co-incubating with USPIO for 24 hours. USPIO particles were mainly located in lysosomes. As demonstrated by trypan-blue staining, there was no significant deference in viability between labeled and unlabeled groups ( P > 0. 05 ). All dual-labeled transplanted BMSCs showed a significant decreasing signal on MRI, and the signal intensity changes had no significant difference over 4 weeks ( P= 0. 66 ). In vitro cell tracing with fluorescence imaging of isolated heart from F344 rats was successful, while in vivo cell tracing with fluorescence imaging failed. Prussian blue staining showed that USPIO distributed near the infracted myocardium, corresponding with the fluorescence imaging. Conclusion MRI can be used to trace the dual-labeled BMSCs transplanted into F344 rat hearts in vivo, while fluorescence imaging and pathological fluorescence imaging can trace the transplanted cells in vitro.关键词
红色荧光蛋白/超小超顺磁性氧化铁/骨髓间充质干细胞/荧光成像/磁共振成像Key words
red fluorescence protein/ultrasmall superparamagnetic iron oxide/bone mesenchymal stem cells/fluorescence imaging/magnetic resonance imaging分类
医药卫生引用本文复制引用
曹剑,王怡宁,石新琳,马国涛,孔令燕,薛华丹,雷晶,何泳蓝,金征宇..双模态标记F344大鼠骨髓间充质干细胞移植入大鼠心脏后离体及在体影像示踪[J].中国医学科学院学报,2012,34(5):474-479,6.基金项目
国家自然科学基金(81000629) （81000629）
