超顺磁性氧化铁标记骨髓间充质干细胞的体外标准化实验

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背景:超顺磁性氧化铁颗粒标记成像可在体观察标记细胞移植后情况,但结合临床细胞治疗剂量,其标记浓度及标记时间对细胞标记率、标记活性及标记后成像效果的综合影响还需进一步分析。目的:筛选超顺磁性氧化铁体外标记大鼠骨髓间充质干细胞的标准化剂量,拟为体内成像提供最佳标记“浓度-时间”。设计、时间及地点:细胞学体外对照观察,于2008-02/10在山西医科大学完成。材料:清洁级Wistar大鼠10只,由山西医科大学动物实验中心提供。Resovist中含超顺磁性氧化铁铁颗粒28g/L,为德国Schering公司产品。方法:全骨髓法分离培养大鼠骨髓间充质干细胞。取Resovist,分别配制含终浓度为0,25,50,75,100,150mg/L超顺磁性氧化铁的培养基,加入P3代骨髓间充质干细胞中进行孵育标记。主要观察指标:普鲁士蓝法检测细胞标记率,锥虫蓝拒染法检测标记细胞活性,观察标记细胞体外磁共振成像情况。结果:细胞标记率达100%,随着标记时间的延长,胞质内蓝染颗粒逐渐减少。标记1d和3d时,与0mg/L超顺磁性氧化铁组比较,25,50mg/L组锥虫蓝拒染率无明显差异(F=0.28~3.15,P>0.05);与25mg/L超顺磁性氧化铁组比较,50mg/L组锥虫蓝拒染率无明显差异(F=0.28~0.79,P>0.05)。体外磁共振成像示标记3d时,标记浓度为50mg/L组的T2WI及T2*WI信号降低最明显。结论:超顺磁性氧化铁“50mg/L-3d”体外标记效率高,对细胞活性无影响,且磁共振成像信号降低最明显,为最适“浓度-时间”组合。 BACKGROUND: Superparamagnetic iron oxide particles labeled imaging can be observed in vivo labeled cells after transplantation, but combined with clinical cell dose, the concentration of the marker and the labeling time on the cell labeling rate, labeling activity and post-labeling imaging effect combined effect further analysis. OBJECTIVE: To screen standardized doses of rat bone marrow-derived mesenchymal stem cells labeled with superparamagnetic iron oxide in vitro and to provide the best marker “concentration-time” for in vivo imaging. DESIGN, TIME AND SETTING: The cytology in vitro control study was performed at Shanxi Medical University on February 02, 2008. MATERIALS: Ten clean Wistar rats were provided by Animal Experimental Center of Shanxi Medical University. Resovist contains superparamagnetic iron oxide particles 28g / L, for the German company Schering products. Methods: Bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow. Take Resovist, were prepared with the final concentration of 0,25,50,75,100,150 mg / L superparamagnetic iron oxide medium, adding P3 generation of bone marrow mesenchymal stem cells for incubation markers. MAIN OUTCOME MEASURES: Prussian blue method was used to detect the cell labeling rate, trypan blue exclusion method to detect the labeled cell activity, and to observe the labeled cells in vitro magnetic resonance imaging. Results: The cell labeling rate reached 100%. With the extension of labeling time, the cytoplasm blue staining particles decreased gradually. Compared with 0 mg / L superparamagnetic iron oxide group, there was no significant difference (P = 0.28 ~ 3.15, P> 0.05) of trypan blue exclusion in 25 and 50 mg / Compared with the paramagnetic iron oxide group, there was no significant difference in the trypan blue exclusion rate in 50 mg / L group (F = 0.28-0.79, P> 0.05). In vitro magnetic resonance imaging marked 3d, T2WI marked marked concentration of 50mg / L group and T2 * WI signal decreased the most obvious. CONCLUSION: Superparamagnetic iron oxide “50mg / L-3d ” has high in vitro labeling efficiency, has no effect on cell activity, and the most obvious reduction of magnetic resonance imaging signal is the optimal “concentration-time” combination.
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