心脏MRI在体示踪移植细胞的可行性研究

何庚戌; 要 彤; 张 浩; 胡盛寿; 宋 鹏; 张晓玲

中国临床医学影像杂志 ›› 2009, Vol. 20 ›› Issue (6) : 418-422.

论著

心脏MRI在体示踪移植细胞的可行性研究

何庚戌1，要彤2，张浩3，胡盛寿3，宋鹏3，张晓玲3

作者信息 +

Study of the feasibility of a novel, clinical applicable cell labeling method to trace the transplanted cells in vivo

HE Geng-xu1, YAO Tong2, ZHANG Hao3, HU Sheng-shou3, SONG Peng3, ZHANG Xiao-ling3

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文章历史 +

摘要

目的：寻找一种能够对移植细胞进行在体示踪的标记方法，为细胞移植的动物和临床实验中细胞的存留、迁移提供重要手段。方法：从中华小型猪髂骨处抽取骨髓，体外培养扩增MSCs。将SPIO和MSCs共同孵育培养36h。普鲁士蓝染色评价细胞的标记效率；通过MTT比色实验评价SPIO对细胞生长能力的影响；台盼蓝染色检验标记后细胞的活性；使用Costar Transwell方法评价铁离子对细胞迁移能力的影响；用细胞分化诱导液培养标记后的细胞评价其向成脂肪细胞和成骨细胞的分化能力。在体内实验中将SPIO标记或未标记的自体MSCs注射到心肌或冠状动脉内，通过心脏磁共振检查对移植细胞进行在体示踪观察。在细胞移植后不同时期取材动物心脏切片进行普鲁士蓝染色观察移植细胞。结果：MSCs经铁离子标记后普鲁士蓝染色阳性率在98%以上，可见蓝染颗粒位于细胞浆内，标记细胞电镜切片观察可见高密度铁颗粒位于细胞浆内。MTT比色实验发现随着培养液中SPIO浓度的增加细胞增殖能力没有明显改变；台盼蓝染色显示标记后98％的细胞保持活性；细胞经SPIO标记后仍可保持原有的细胞形态，可继续培养、传代。细胞迁移试验发现细胞标记后对SDF-1和VEGF诱导的迁移能力没有明显的减低。铁离子标记后细胞仍可向成脂肪细胞和成骨细胞分化。注射到心肌或冠状动脉内的SPIO标记的MSCs可通过心脏磁共振检查进行在体示踪，动态观察显示SPIO标记细胞在磁共振影像上表现为低密度灶影或信号缺失，并且在移植后4周仍可显影。病理学检查可以看到移植细胞呈普鲁士蓝染色阳性，并和影像学有很好的一致性。结论：临床使用的SPIO磁共振对比剂可以安全、有效的标记MSCs。利用心脏磁共振对移植的标记细胞可以实现移植细胞的在体示踪观察。此标记方法结合心脏磁共振技术为移植细胞的在体示踪和细胞存留、迁移的研究提供了良好的手段。

Abstract

Objective: To find a new labeling agent to trace and visualize the transplanted cells in vivo. Methods: Bone marrow MSCs were incubated with SPIO for 36 hours. The labeling efficiency was tested through Prussian Blue staining, the growth ability was evaluated through MTT, and cells viability was tested through Trypan Blue rejection method, the migratory ability was assessed with Costar Transwell plates, the differentiation of labeled cells to adipogenic and osteogenic cells was induces with induction medium. The labeled or unlabeled cells were transplanted into the myocardium or coronary artery. MRI was carried out immediately and 1～4 weeks respectively. After MRI the heart was excised, the segment in which injections performed were thin cut and stained with hematoxylin-eosin and Prussian Blue staining. Results: There were intracytoplasmic blue particles in nearly every cell in Prussian Blue staining. SPIO had no poison effect on the cell growth and proliferation. The viability of cells was more than 98%. The migratory ability was not affected. The labeled MSCs could still be differentiated from adipogenic and osteogenic cells after being incubated in the inductive medium. The labeled cells could be detected through MRI and confirmed by pathology. After 4 weeks the injected labeled cells could still be detected through MRI. The pathology showed the injected cells could survive in the MI area, and parallel in the same direction. Conclusion: The cells could be efficiently and safely labeled with SPIO and the labeled cells could be reliably detected by MRI in vivo.

导出引用

何庚戌;要彤;张浩;胡盛寿;宋鹏;张晓玲. 心脏MRI在体示踪移植细胞的可行性研究[J]. 中国临床医学影像杂志. 2009, 20(6): 418-422

HE Geng-xu;YAO Tong;ZHANG Hao;HU Sheng-shou;SONG Peng;ZHANG Xiao-ling. Study of the feasibility of a novel, clinical applicable cell labeling method to trace the transplanted cells in vivo[J]. Journal of China Clinic Medical Imaging. 2009, 20(6): 418-422

中图分类号： Ｒ654.2 Ｒ445.2