PLL-Ferumoxytol标记小鼠胰岛β细胞系MIN6的生物活性研究  

Characterization of biophysical properties of isletβcell line MIN6 labeling with PLL-Ferumoxytol

作　　者：Silvany Simone Sanches TAVARES 张淼 祝晓阳 顾宁[1] Silvany Simone Sanches TAVARES;ZHANG Miao;ZHU Xiaoyang;GU Ning(Jiangsu Key Laboratory for Biomaterials and Devices,State Key Laboratory of Bioelectronics,School of Biological Science and Medical Engineering,Southeast University,Nanjing 210009,China)

机构地区：[1]东南大学生物科学与医学工程学院,生物电子学国家重点实验室,江苏省生物材料与器件重点实验室,江苏南京210009

出　　处：《东南大学学报（医学版）》2020年第6期697-704,共8页Journal of Southeast University(Medical Science Edition)

基　　金：国家重点研究开发项目(2017YFA0104302);国家自然科学基金资助项目(61821002,51832001);江苏省科学委员会青年基金资助项目(BK20180374)。

摘　　要：目的:将多聚赖氨酸(PLL)与本课题组自行制备的超顺磁性氧化铁纳米粒子Ferumoxytol结合,利用PLL修饰的Ferumoxytol标记小鼠胰岛β细胞系MIN6细胞,初步探索其标记MIN6细胞的可行性、标记效率以及生物安全性等。方法:用Ferumoxytol及PLL修饰的Ferumoxytol与小鼠胰岛β细胞系MIN6细胞共孵育标记细胞,采用细胞增殖检测试剂盒(CCK-8)实验检验MIN6细胞的活力与增殖情况,利用肌动蛋白丝染色观测细胞内部细胞骨架的形貌,普鲁士蓝染色和透射电镜观察被标记的细胞内铁颗粒的分布情况,使用葡萄糖刺激胰岛素释放试验(GSIS)对标记后MIN6细胞的胰岛素合成及分泌功能进行检验和评估。结果:标记后细胞与对照组的形貌以及细胞骨架特性均无显著差异,且均维持了较高活性。添加200μg·ml^-1 PLL-Ferumoxytol共培养24 h可以显著促进细胞增殖。PLL-Ferumoxytol标记组细胞胞质内呈现蓝色铁颗粒,透射电镜结果进一步验证了铁颗粒的分布情况。此外,添加纳米粒子对MIN6细胞胰岛素合成与释放功能无显著影响。结论:本研究小组自行制备的PLL-Ferumoxytol具有良好的生物相容性,能够有效标记小鼠MIN6细胞,对细胞的活性和功能无显著性影响,可以进一步用作胰岛细胞移植的磁标记物并应用于MRI活体示踪。Objective:The superparamagnetic iron oxide nanoparticles of Ferumoxytol were synthesized with poly-L-lysine(PLL)and used to label mouse isletβcell line MIN6.The feasibility,labeling efficiency and biosafety of PLL-Ferumoxytol were preliminarily explored.Methods:MIN6 cells were cocultured with Ferumoxytol and PLL-Ferumoxytol.Cell counting kit-8(CCK8)was used to test the viability and proliferation of MIN6 cells.Actin filament staining was performed to characterize the cytoskeleton morphology.Prussian blue staining and transmission electron microscopy(TEM)were used to detect and characterize the distribution of iron particles in labeled cells.Glucose stimulated insulin secretion(GSIS)test was performed to evaluate the function of insulin synethesis and secretion of labeled MIN6 cells.Results:No significant difference was detected in cell morphology and cytoskeleton properties between the labled and control groups.High viablilies of cells in all groups were maintained.Moreover,after incubating with 200μg·ml^-1 PLL-Ferumoxytol for 24 hours,cell proliferation was significantly improved.PLL-Ferumoxytol labeled cells had blue-stained iron particles in the cytoplasm,and TEM results further verified the distribution of these particles in cells.In addition,coculturing the cells with nanoparticles had no significant effect on the insulin synthesizing and secreting functions of MIN6 cells.Conclusion:PLL-Ferumoxytol has good biocompatibility,and can be used to effectively label MIN6 cells with no significant effect on viablity and function of the cells.Therefore,we conclude that as an effective magnetic marker,PLL-Ferumoxytol can be applied for in vivo MRI tracking in islet transplantationin the future.

关 键 词：FERUMOXYTOL 多聚赖氨酸-Ferumoxytol MIN6细胞 胰岛移植 小鼠 

分 类 号：R-33[医药卫生]