Influence on bionomics of endothelial progenitor cells labeling with magnetic nanoparticles / 中华放射学杂志

ABSTRACT

Objective To explore the influence of home synthesize magnetic iron oxide (called Fe2O3-PLL) labeling on peripheral blood endothelial progenitor cells (EPCs) bionomics to provide experimental foundation for MR imaging ex and in vivo. Methods Fe2O3 was incubated with PLL for 2 hours to obtain a complex of Fe2O3-PLL. Rabbit peripheral blood mononuclear cells were isolated and EPCs were selected by adherence method. Fe2O3-PLL was used to label EPCs. Prussian blue stain and electron microscope was used for showing intracellular iron. MTT assay was assessed to evaluate the difference of growth curve between unlabeled and labeled with 25 mg/L Fe2O3-PLL. Flow cytometry was performed to analyze cell cycle, cell apoptosis and the expression of surface markers of labeled and unlabeled cells. Expressions of Enos, KDR and Vwf at Mrna levels among unlabeled and labeled EPCs were detected by real-time polymerase chain reaction. Calcium ion channel and membrane fluidity were observed and analyzed by laser confocal microscopy. Statistical analyses were used with ANOVA and t test. Results Almost 100% cells were labeled by Fe2O3-PLL, iron-containing vesicles were intracytoplasma. There was no statistical difference in cells growth curve, cell life cycle [(93.74±3.52)% ,(94.57±3.66)% ] and cell apoptosis rate(12. 89±1.81) %, (11.67±1.18) %) between labeling with Fe2O3-PLL at a concentration of 25 mg/L and unlabeled cells (t = 0. 283, P > O. 05 ; t = 0. 977, P > 0. 05). There was also no statistical difference in relative amount of Enos, KDR and Vwf at Mrna levels and the expression of sudace phenotypic markers (CD34, CD106, CD146 and KDR) between two groups (P > 0. 05). In addition,Labeling had little influence on calcium ion channel and didn't significantly alter cell membrane fluidity.Conclusions The rabbit peripberal blood EPCs can be effective labeled with Fe2O3-PLL and without significant influence on cells bionomics at a low concentration of 25 mg/L. Almost every cell can be labeled and the labeled cells can be used further.