Shen Jun, Zhong Xiao-Mei, Duan Xiao-Hui, Cheng Li-Na, Hong Guo-bing, Bi Xiao-Bing, Liu Yu
Department of Radiology, The Second Affiliated Hospital, Sun Yat-sen University, 107 Yanjiang Road West, Guangzhou 510120, Guangdong, China.
Acad Radiol. 2009 Sep;16(9):1142-54. doi: 10.1016/j.acra.2009.03.016.
RATIONALE AND OBJECTIVES: In vivo tracking cells using gadolinium-based contrast agents have the important advantage of providing a positive contrast on T1-weighted images, which is less likely to be confused with artifacts because of postoperative local signal voids such as metal, hemorrhage, or air. The aim of this study is to paramagnetically and fluorescently label marrow with dual agents (gadolinium-diethylene triamine penta-acetic acid [Gd-DTPA] and PEI-FluoR) and track them after transplantation into spinal cord injury (SCI) with magnetic resonance imaging (MRI). MATERIALS AND METHODS: Marrow mesenchymal stem cells (MSCs) from Sprague-Dawley rats were incubated with PEI-FluoR (rhodamine-conjugated PEI-FluoR) and Gd-DTPA complex for labeling. After labeling, cellular viability, proliferation, and apoptosis were evaluated. T1 value and longevity of intracellular Gd-DTPA retention were measured on a 1.5 T MRI scanner. Thirty-six SCI rats were implanted with labeled and unlabeled MSCs and phosphate-buffered saline. Then, serial MRI and Basso-Beattie-Bresnehan (BBB) locomotor tests were performed and correlated with fluorescent microscopy. The relative signal intensity (RSL) of the engraftment in relation to normal cord was measured and the linear mixed model followed by post-hoc Bonferroni test was used to identify significant differences in RSL as well as BBB score. RESULTS: MSCs could be paramagnetically and fluorescently labeled by the dual agents. The labeling did not influence the cellular viability, proliferation, and apoptosis. The longevity of Gd-DTPA retention in labeled MSCs was up to 21 days. The distribution and migration of labeled MSCs in SCI lesions could be tracked until 7 days after implantation on MRI. The relative signal intensities of SCI rats treated with labeled cells at 1 day and 3 days (1.34 +/- 0.02, 1.27 +/- 0.03) were significantly higher than rats treated with unlabeled cells (0.94 +/- 0.01, 0.99 +/- 0.02) and phosphate-buffered saline (0.91 +/- 0.01, 0.95 +/- 0.01) (P < .05). Rats treated with labeled MSCs or unlabeled MSCs achieved significantly higher BBB scores than controls at 14, 21, 28, and 35 days after injury (P < .05). CONCLUSIONS: Labeling MSCs with the dual agents may enable cellular MRI and tracking in experimental spinal cord injury.
原理与目的：使用基于钆的造影剂在体内追踪细胞具有重要优势，即能在T1加权图像上提供阳性对比，不太可能因术后局部信号缺失（如金属、出血或空气）产生的伪影而混淆。本研究的目的是用双试剂（钆 - 二乙烯三胺五乙酸[Gd - DTPA]和PEI - FluoR）对骨髓进行顺磁性和荧光标记，并在将其移植到脊髓损伤（SCI）后用磁共振成像（MRI）进行追踪。 材料与方法：将来自Sprague - Dawley大鼠的骨髓间充质干细胞（MSCs）与PEI - FluoR（罗丹明偶联的PEI - FluoR）和Gd - DTPA复合物一起孵育进行标记。标记后，评估细胞活力、增殖和凋亡情况。在1.5T MRI扫描仪上测量细胞内Gd - DTPA保留的T1值和持续时间。36只SCI大鼠分别植入标记和未标记的MSCs以及磷酸盐缓冲盐水。然后，进行系列MRI和Basso - Beattie - Bresnehan（BBB）运动测试，并与荧光显微镜检查结果相关联。测量植入物相对于正常脊髓的相对信号强度（RSL），并使用线性混合模型及事后Bonferroni检验来确定RSL和BBB评分的显著差异。 结果：MSCs可用双试剂进行顺磁性和荧光标记。标记不影响细胞活力、增殖和凋亡。标记的MSCs中Gd - DTPA保留的持续时间长达21天。标记的MSCs在SCI损伤中的分布和迁移在MRI上可追踪至植入后7天。在第1天和第3天，用标记细胞治疗的SCI大鼠的相对信号强度（1.34±0.02，1.27±0.03）显著高于用未标记细胞（0.94±0.01，0.99±0.02）和磷酸盐缓冲盐水（0.91±0.01，0.95±0.01）治疗的大鼠（P <.05）。在损伤后14、21、28和35天，用标记或未标记MSCs治疗的大鼠的BBB评分显著高于对照组（P <.05）。 结论：用双试剂标记MSCs可能实现实验性脊髓损伤中的细胞MRI成像和追踪。
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