摘要
目的磁粒子标记骨髓间充质干细胞,同种异体移植入急性肝损伤大鼠脾脏,应用1·5TMR仪进行活体成像,为干细胞移植并无创示踪提供实验依据。方法制备Fe2O3-多聚左旋赖氨酸(PLL),标记分离纯化后的大鼠骨髓间充质干细胞(bonemesenchymalstemcells,BMSCs),普鲁士蓝染色显示细胞内铁。12只急性肝损伤大鼠分成实验和对照两组,将标记细胞(实验组,n=6)和未标记细胞(对照组,n=6)同种异体移植入大鼠脾脏,在移植前、移植后3h及3、7、14d应用MR的T2W对肝脏进行活体成像,测量肝脏信噪比(signal-to-noiseratio,SNR),并与肝脏组织切片普鲁士蓝染色对照。结果BMSCs的Fe2O3-PLL标记率近100%,普鲁士蓝染色显示蓝色铁颗粒位于BMSCs胞质内。实验组和对照组注射前、注射后3h及3、7、14d肝脏的SNR分别为19·53±2·30,3·28±1·06,7·34±2·10,10·25±3·96,15·50±3·73;20·20±4·35,21·20±4·43,19·13±2·80,21·43±5·45,19·07±4·80。与注射前比较,实验组3h及3、7d肝脏的SNR下降,差异具有统计学意义(Dunnett检验,t值分别为16·25,12·19,9·29,P值均<0·05),14d时SNR虽仍较低,但差异无统计学意义(Dunnett检验,t=4·03,P>0·05)。对照组移植后各时间点与移植前比较,SNR改变差异无统计学意义(方差分析,F=0·187,P>0·05)。组织学示普鲁士蓝阳性细胞主要分布于肝小叶中央静脉周围病变区。结论Fe2O3-PLL可以有效标记大鼠BMSCs,临床应用型1·5TMR仪可对磁粒子标记的BMSCs经脾植入后进行活体示踪。
Objective To evaluate the 1.5-T magnetic resonance imaging system to depict and track in vivo of magnetically labeled bone mesenchymal stem cells （BMSCs） in rat liver after intrasplenic transplantation. Methods Rat BMSCs were isolated, purified, expanded and then incubated with home synthesized Fe2O3-PLL. Prussian blue stain was performed for showing intracellular irons. Acute liver damage was induced by subcutaneous injection of carbon tetrachloride in 12 recipient rats. The cells allotransplantation was performed by intrasplenic injection with magnetically labeled （experimental group, n=6） or unlabeled BMSCs （control group, n=6）. Serial MRI were performed before and 3 hours, 3, 7, 14 days after transplantation. Signal-to-noise ratio （SNR） of liver on T2 ＊-weighted MR imaging obtained before and after injection were measured and compared. MR imaging findings were compared histologically with histology. Results Rats BMSCs could be effectively labeled and the labeling efficiency was almost 100%. Iron-containing intracytoplasmatic vesicles could be observed clearly with prussian blue staining. SNR of rat livers in experimental group and control group before and 3 hours, 3, 7, 14 days after transplantation were 19.53 ± 2. 30,3.28 ± 1.06,7.34 ± 2.10,10. 25 ± 3.96,15.50 ± 3.73 ; 20. 20 ± 4. 35, 21.20 ±4. 43,19.13 ±2. 80,21.43 ±5.45,19.07 ±4. 80, respectively. SNR decreased significantly in the experimental group liver 3 hours, 3, 7 days after injection of BMSCs （ Dunnett test ,P 〈 O. 05 ） , and returned gradually at approximately day 14 to that of rat livers before injection （ Dunnett test, P 〉 0. 05 ）. In control group, SNR demonstrated no significant differences among different time points （ ANOVA, P 〉 0. 05 ）. Results of histological analysis confirmed homing of labeled BMSCs in liver, primarily distributing in areas around centrolobular vein. Conclusion The BMSCs can be effectively labeled with Fe2O3-PLL. 1.5-T MR imaging can monitor in vivo of magnetically labeled BMSCs in liver after intrasplenic transplantation. It potentially opens a new area of investigation for delivering stem cells.
出处
《中华放射学杂志》
CAS
CSCD
北大核心
2006年第2期127-132,共6页
Chinese Journal of Radiology
基金
国家自然科学基金资助项目(30400116)
高等学校博士学科点专项科研基金资助项目(20040286037)
江苏省自然科学基金资助项目(BK2004068)
东南大学国家自然科学基金预研项目(XJ0490170)
关键词
间充质干细胞
磁共振成像
标记
移植
肝脏
Mesenchymal stem cells
Magnetic resonance imaging
Labeling
Transplantation
Liver
