磁纳米颗粒标记脂肪干细胞向软骨分化及体外MRI示踪的实验研究  

In Vitro MRI Monitoring Magnetic Iron Oxide Particles(MIOP) Labeling Adipose Derived Stem Cells Differentiation into Chondrocytes

作　　者：祝加学[1] 陆雄伟[1] 胡小鹏[1] 顾剑华[1] 王健[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院骨科,上海201999

出　　处：《现代生物医学进展》2016年第12期2209-2212,共4页Progress in Modern Biomedicine

基　　金：上海交通大学医学院基金项目(13XJ10055);上海市卫生局科研基金项目(20134195)

摘　　要：目的:探讨磁纳米颗粒(magnetic iron oxide particles,MIOP)体外标记脂肪间充质干细胞(ASCs)向软骨分化及MRI示踪的可行性。方法:从小鼠脂肪组织中分离培养、扩增脂肪间充质干细胞(ASCs),流式鉴定细胞表型后,分别采用不同浓度(25μg/mL,50μg/mL)的MIOP标记ASCs并向软骨细胞诱导分化。普鲁士蓝染色和透射电镜(TEM)鉴定细胞内磁纳米铁颗粒分布情况,应用3.0T MRI体外检测标记软骨细胞MRI信号。结果:从脂肪组织中可以分离获得大量高表达CD90、CD105、Sca-1的ASCs,不同浓度(25μg/mL,50μg/mL)的MIOP与ASCs共同孵育24小时后,普鲁士蓝染色发现ASCs随MIOP浓度的增加,蓝染程度逐渐加深且标记的ASCs可以向软骨细胞分化;TEM证实细胞内分布大量的黑色纳米铁颗粒。体外MRI T2序列证实随着MIOP浓度(25μg/mL,50μg/mL)的增加MRI信号值逐渐减低且具有统计学差异(P<0.05)。结论:MIOP可以标记ASCs向软骨分化,体外应用MRI可以对其进行示踪。Objective： To explore the feasibility and effects of labeling Adipose derived stem cells（ASCs） differentiation into chondrocytes with magnetic iron oxide particles（MIOP）, and to investigate the possibility of in vitro MR monitoring. Methods： Adipose derived stem cells（ASCs） were isolated, expanded from mouse fat tissues, P3 ASCs were tested by Flow cytometry and labeled with different concentration of MIOP（25 μg/mL, 50 μg/mL） and differentiated into chondrocytes. Prussian blue staining and Transmission Electron Microscope（TEM） were performed to show the intracellular iron particles. In vitro 3.0 MRI were used to test the signal intensity of labeled cells. Results： ASCs were successfully cultured and high expression CD90, CD105, Sca-1. After ASCs were cultured with different concentration of MIOP（25 μg/mL, 50 μg/mL） about 24 hours. With the MIOP concentration increase from 25 μg/mL to50 μg/mL, the dyeing degree of labeling cells stained by Prussion blue staining deepened and the labeled ASCs could differentiate into chondrocytes. TEM indicated that a lot of black iron particles accumulated in the cells. In vitro 3.0 MRI results show that T2 intensity of labeling cells were inversely correlated with the MIOP（25 μg/mL, 50 μg/mL）（P〈0.05）compared with the control group. Conclusions：Adipose derived stem cells could be labeled effectively with MIOP and differentiate into chondrocytes, and in vitro MRI could be used to monitor these labeled cells.

关 键 词：软骨分化 脂肪间充质干细胞(ASCs) 磁纳米铁颗粒(MIOP) MRI示踪 

分 类 号：R95-3[医药卫生—药学] Q64[生物学—生物物理学]