磁标记兔脂肪间充质干细胞生物学特性及体外MRI - 第三军医大学学报.DOC

兔脂肪间充质干细胞的培养鉴定及体外磁标记MR成像( 谢光友 杨海涛 吕富荣 鲁文力 盛波 肖智博 吕发金 欧阳羽 (重庆医科大学附属第一医院放射科 重庆 400016) 【摘要】目的 研究兔脂肪间充质干细胞（ADSCs）的培养鉴定方法，联合应用超顺磁性氧化铁（SPIO）和多聚赖氨酸（PLL）行体外磁标记细胞，并探讨磁标记示踪的可行性。方法 分离、纯化并培养兔ADSCs，流式细胞术鉴定细胞表面抗原CD90、CD44和CD34。应用SPIO-PLL标记细胞，普鲁士蓝染色和透射电镜检测胞内铁粒子。体外3.0T MR分别对1×107个未标记细胞，经25、50、75μg/ml SPIO-PLL标记的1×107个细胞；1×107个ADSCs（标记1d）、1×107个（标记3d）和5×106个（标记1d）））The culture and identification of rabbit adipose stem cells and its magnetic resonance imaging in vitro Xie Guangyou , Yang Haitao, Lv Furong, Lu Wenli,Sheng Bo, Xiao Zhibo, Lv Fajin , OuYang Yu (Department of Radiology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China) 【Abstract】Objective:To study the method of culture and identification of rabbit adipose stem cells, labeling the cells by superparamagnetic iron oxide particles(SPIO) combined with poly-l-lysine(PLL) in virto,and to discuss the feasibility of SPIO -PLL to trace ADSCs.Methods:ADSCs were obtained by means of isolation, purification and culture,and the cell surface antigens CD44, CD90 and CD34 were identified through flow cytometry(FCM). The intracellular iron particles were detected by Prussian blue staining and transmission electron microscopy (TEM) .1×107 ADSCs unlabeled and labeled with 25, 50, 75μg/ml SPIO-PLL respectively were performed with 3.0 T MR scanning in vitro ,meanwhile,1×107ADSCs (labeled 1d), 1×107 (labeled 3d) and 5×106((labeled 1d) were also scanned, including T1WI, T2WI and T2*WI arrays, the signal intensity (SI) and relaxation time of each tube were measured. Result:The third generation ADSCs has a high degree of purity,arranged regularly,and presented whirlpool-like.The expression rate of CD44 and CD90 were 98.7%, 99.2% respectively, but CD34 expressed negatively.The results of Prussian blue staining and TEM showed that particles existed in each cytoplasm,and the rate of labeling was nearly 100%. T2*WI and T2WI demonstrated a more obvious decrease in SI than that of T1WI with the increasing concentrat