超顺磁性纳米铁标记大鼠骨髓基质干细胞的实验研究

Studies on rat mesenchymal stem cells labeling with superparamagnetic iron oxide

目的研究超顺磁性纳米铁粒子(superparamagnetic iron nanoparticle,SPIO)标记骨髓基质干细胞(bone mar-row stem cells,BMSCs)对其存活、增殖能力的影响,确定最佳标记浓度。方法分离、纯化4周龄SD大鼠的BM-SCs,分别用15、25、50 mg/L浓度的SPIO标记1×106个BMSCs分别作为实验组1、实验组2、实验组3,未做标记的BMSCs为对照组。用普鲁士蓝染色法和透射电镜鉴定SPIO-BMSCs的效率,台蓝染色法观察SPIO-BMSCs的存活,MTT比色法检测SPIO-BMSCs增殖活性。结果普鲁士蓝染色显示,随SPIO浓度的升高,标记的细胞染色程度逐渐加深。透射电镜检查显示细胞内含致密铁颗粒。实验组1的细胞增殖性与对照组差异无统计学意义(P>0.05),实验组2和实验组3与对照组相比,死亡细胞增多,细胞增殖性受到明显抑制(P<0.05)。结论培养基SPIO浓度在15 mg/L时,SPIO对BMSCs标记效率较高,且不影响其存活、增殖。

Objective To study the effect of superparamagnetic iron nanoparticle（SPIO） labeling on the ability of bone marrow stem cells（BMSCs） to survive and proliferate,and to investigate the optimal concentration of SPIO.Methods BMSCs isolated from SD rats（4 weeks old） were expanded,and then l ×106 cells were labeled with the concentration of 15,25 mg/L and 50 mg/L SPIO and were recorded as experimental group 1,experimental group 2 and experimental group 3 respectively.Prussian blue staining and transmission electron microscope（TEM） were performed to investigate the efficiency of SPIO labeling.Taiwan blue staining was used to observe the survival of SPIO-BMSCs,and MTT col orimetric method to detect their proliferative activity.Results Dyeing degree of labeled cells stained by Prussian blue was increased when the concentration of SPIO increased from 15 mg/L to 50 mg/L.Transmission electron microscope indicated that iron particles were accumulated in the lysosomes in the cytoplasm.No significant difference of the abili ties to survive and proliferate was noticed between cells in the experimental group 1 and the control group（P 0.05）.Dead cells were increased and proliferative ability was decreased in cells of the experimental group 2 and experimental group 3,compared to the control group（P 0.05）.Conclusion The labeling efficiency is optimal and the ability of BMSCs to survive and proliferate is not affected when the concentration of SPIO is 15 mg/L.