荧光磁性纳米粒子体外标记骨髓间充质干细胞研究

Study of fluorescent supermagnetic nanoparticles labeled bone marrow mesenchymal stem cells in vitro

目的:探索高效体外示踪骨髓间充质干细胞体系的方法。方法:采用贴壁培养法分离、纯化大鼠骨髓间充质干细胞,用流式细胞仪检测骨髓间充质干细胞CD29、CD90和CD45表达情况,用诱导剂诱导干细胞检测骨髓间充质干细胞的分化能力,采用普鲁士蓝染色,用激光共聚焦显微镜、磁共振成像仪观察荧光磁性纳米粒子标记的干细胞体外成像效果。结果:流式细胞仪检测CD29、CD90和CD45阳性表达率分别为84.69%、90.28%和0.72%;碱性磷酸酶染色和油红O染色结果均表明骨髓间充质干细胞成功分化为成骨细胞和脂肪细胞;普鲁士蓝染色、激光共聚焦显微镜成像和磁共振成像表明荧光磁性纳米粒子标记的骨髓间充质干细胞具有很好的成像效果。结论:荧光磁性纳米粒子作为示踪剂可以在体外标记骨髓间充质干细胞,为干细胞研究提供新思路。

Objective： To establish a system for efficiently tracking bone marrow mesenchymal stem cells（BMSCs） in vitro.Methods： BMSCs were harvested and purified from rats by adherent culture and then BMSCs expression of CD29,CD90 and CD45 were detected by using flow cytometry.To evaluate their ability to differentiate into multi-lineage cells,BMSCs were cultured in different induced media.Prussian blue staining,laser scanning confocal microscope,magnetic resonance imaging were used to observe labeled BMSCs in vitro after co-culturing with FMNPs.Results： FACS analysis showed that these cells were positive for markers CD29,CD90,CD45 as 84.69%,90.28% and 0.72%.ALP staining and oil red O staining indicated that BMSCs were inducted into osteoblasts and adipose cells.Prussian blue staining,laser scanning confocal and magnetic resonance imaging showed that the prepared FMNPs labeled BMSCs had high quality images.Conclusion： As an important stem cell tracer,FMNPs could label BMSCs and provided a new method for stem cell research.