目的 探索简单、无细胞外铁产生的超低微浓度菲立磁-硫酸鱼精蛋白标记骨髓间充质干细胞(MSCs)方法.方法 贴壁法培养大鼠MSCs.待3代细胞汇合至80％～90％时,更换无血清培养液,根据菲立磁和硫酸鱼精蛋白的不同浓度分为4组:A组[(7.50:1.00)μg/ml]、B组[(10.00:1.20)μg/ml];C组[(15.00:1.80)μl/ml]和空白对照组,加入培养液,混匀,5％ CO2孵育15 min,补加血清后孵育至次日.检测细胞标记率、细胞内外铁、细胞活力和标记细胞MR信号.结果 B组可有效标记大鼠MSCs,普鲁士蓝染色阳性率100％,无细胞外铁产生,铁颗粒分布于溶酶体内.4组间台盼蓝拒染率差异无统计学意义(P＞0.05)；体外MR GRE T2* WI序列可检测到1×104个标记细胞.结论 使用超低微浓度菲立磁10.00μg/ml与鱼精蛋白1.20 μg/ml可有效标记大鼠MSCs,体外MR可检测到1×104个标记细胞.%Objective To explore a simple, no extracellular iron produced labeling method of ultra-low micro concentration of feridex-protamine complexes labeling mesenchymal stem cells (MSCs). Methods MSCs were incubated by adherence method. When the 3th passage cells reached 80%-90% confluence, the old media with serum free DMEM was placed. According to different concentrations of feridex and protamine, 4 groups were classified, I. E. Group A ([7. 50 : 1. 00]μg/ml) , B ([10. 00 : l. 20]μg/ml), C ([15. 00 : l. 80]μl/ml) and blank control group. Nutrient solution were added in the feridex-protamine complexes and blended. After incubating 15 min at 37℃ , 5% CO2, serum was added and the labeling procedure was further continued over night. The cell labeling efficiency, intracellular and extracellular iron, MR signal of labeled cells were detected. Results MSCs could be efficiently labeled in Group B. The Prussian blue staining positive rate was almost 100% , there was no extracellular iron. Iron distributed evenly in the lysosomes. There was no statistical difference among the 4 groups (P>0. 05). A total of 1 × 104 labeled cells could be detected by in vitro MR GRE T2 WI sequence. Conclusion MSCs could be efficiently labeled in the presence of ultra-low micro concentration feridex-protamine complexes [(10. 00 : l. 20)μg/ml], 1 × 104 labeled cells could be detected by in vitro MR with the mentioned method.
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