摘要
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 labeled imaging can monitor labeled cell in vivo. However, combined with clinical cell therapy dose, effects of different labeled concentration and labeled time on the labeling cell efficiency, the labeling cell viability and magnetic resonance imaging of labeled cells need further research. OBJECTIVE： To explore standard dose of superparamagnetic iron oxide labeled rat bone marrow mesenchynal stem cells （BMSCs） to provide an optimal labeling ＂concentration-time＂ in vivo imaging. DESIGN, TIME AND SETTING： The cytological in vitro controlled study was performed at the Shanxi Medical University from February to October 2008. MATERIALS： Ten clean Wistar rats were supplied by Animal Experimental Center, Shanxi Medical University. Resovist containing 28 g/L superparamagnetic iron oxide was purchased from Schering, Germany. METHODS： Rat BMSCs were isolated and cultured by the whole bone marrow method. Resovist was used to prepare medium, supplemented with 0, 25, 50, 75, 100, 150 mg/L superparamagnetic iron oxide. BMSCs at passage three were incubated and labeled. MAIN OUTCOME MEASURES： Prussian blue method was used to detect cell labeling rate. Trypan blue exclusion test was utilized to label cell activity./n vitro magnetic resonance imaging of labeled cells was observed. RESULTS： The labeling efficiency reached to 100%. With time prolonged, blue-stained particles gradually less in cytoplasm. At 1 and 3 days, compared with the 0 mg/L superparamagnetic iron oxide group, no significant difference in trypan blue exclusion rate in the 25 and 50 mg/L superparamagnetic iron oxide group was detected （F =0.28 3.15, P 〉 0.05）. Compared with the 25 mg/L superparamagnetic iron oxide group, no significant difference in trypan blue exclusion rate in the 50 mg/L superparamagnetic iron oxide group was detected （F=0.28-0.79, P 〉 0.05）. At 3 days of in vitro nuclear magnetic resonance labeling, T2WI and T2＊WI signal significantly reduced in the 50 mg/L superpa
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第19期3633-3639,共7页
Journal of Clinical Rehabilitative Tissue Engineering Research
