多元醇法合成SPIO标记大鼠ADSCs的成神经诱导潜能及体外MR成像  

Labeling of adipose derived stem cells with polyol process of superparamagnetic iron oxide for neural induction of rats and MR imaging in vitro

作　　者：马伟琼[1,2] 谢琦[1] 张鼎旋[1] 汤间仪[1] 雷正贤 杨逸铭[1] 

机构地区：[1]广州医科大学附属广州市第一人民医院广州市南沙中心医院医学影像部,广东广州510180 [2]惠州市中心人民医院放射科,广东惠州516001

出　　处：《中国介入影像与治疗学》2015年第11期686-690,共5页Chinese Journal of Interventional Imaging and Therapy

基　　金：2014广州市南沙区经贸科技与信息化局民生科技项目基金(2014MS01);2014广州市医药卫生科技一般引导项目(20141A010015);2015广州市科技计划项目科学专项(一般项目)基金(1563000424);2015广东省科技计划项目技术开发类基金(2014A020212034)

摘　　要：目的 探讨采用多元醇法合成的SPIO标记SD大鼠脂肪来源干细胞（ADSCs）,进行成神经诱导和体外MRI成像的可行性。方法 分离、纯化、鉴定SD大鼠来源的ADSCs;确定安全有效的标记浓度和时间后,将SPIO与ADSCs共孵育。体外MRI条件下,采用T2-mapping序列观察标记组的标记细胞数、标记持久性,以及标记后标记组与未标记组间信号强度和T2值的差异。ICP-AES检测MRI中标记细胞的单铁含量。结果 12μg/ml、25μg/ml PEG/PEI修饰的SPIO和25μg/ml PEG/PVP修饰的SPIO孵育12h后,可安全有效地对ADSCs进行标记;成神经诱导后,神经标记物Nestin、NSE有明显表达。体外MRI显示标记组的T2WI信号强度和T2值与未标记组差异均有统计学意义（P均〈0.001）;103个细胞量即可引起T2WI信号和T2值的改变;PEG/PEI修饰的SPIO标记持久性维持到20天,PEG/PVP修饰的标记持久性维持到15天。当铁含量为1.56~1.8pg/cell时,标记细胞的T2值与未标记细胞差异无统计学意义（P〉0.05）。结论 多元醇法合成的SPIO可直接对ADSCs进行标记,标记后不仅可向神经方向诱导分化,还可进行体外MRI示踪成像。Objective To explore the feasibility of labeling SD-rat adipose derived stem cells （ADSCs） with superpara- magnetic iron oxide （SPIO） prepared by using polyol method for neural induction and MR imaging in vitro. Methods After isolation, purification and identification, the SD-rat ADSCs were incubated at appropriate concentration and time which had been detected in labeling efficiency and biosecurity repeatedly. In vitro, MRI were performed with T2-mapping sequence for SPIO-labeled cells. Labeled cells undergone continued passage to explore the labeled persistence. Meanwhile, the iron content of labeled cells were assessed by ICP-AES to detect the iron content per cell. Results The ADSCs could be incubated directly, safely and effectively in an appropriate condition： The concentration was 12μg/ml, 25μg/ml with PEG/PEI- SPIO and 25μg/ml with PEG/PVP-SPIO, and both of the time were 12 h. After nerve-induced, neural marker including Nestin and NSE had obvious expression. The MR signal intensity and relaxation time in T2-mapping sequence showed a significant statistical difference between labeled cells and unlabeled cells （all P〈0. 001） 103 labeled cells were detected by MRI. The effective time detected labeled cells by MRI was 20 days with PEG/PEI-SPIO and 15 days with PEG/PVP- SPIO. When the iron content decreased to 1.56--1.8 pg per cell, the relaxation time of labeled cells was similar to unla- beled cells （P〈0.05）. Conclusion The SPIO which is synthesized by an improved polyol method could label ADSCs di- rectly, which appeares to be sensitive neural differentiation and MRI imaging in vitro.

关 键 词：超顺磁性氧化铁 成神经诱导 脂肪干细胞 磁共振成像 体外 

分 类 号：R-332[医药卫生]