Prussian Blue Nanoparticle-Labeled Mesenchymal Stem Cells: Evaluation of Cell Viability, Proliferation, Migration, Differentiation, Cytoskeleton, and Protein Expression In Vitro

Abstract Mesenchymal stem cells (MSCs) have been used for the treatment of various human diseases. To better understand the mechanism of this action and the fate of these cells, magnetic resonance imaging (MRI) has been used for the tracking of transplanted stem cells. Prussian blue nanoparticles (PBNPs) have been demonstrated to have the ability of labeling cells to visualize them as an effective MRI contrast agent. In this study, we aimed to investigate the efficiency and biological effects of labeled MSCs using PBNPs. We first synthesized and characterized the PBNPs. Then, iCELLigence real-time cell analysis system revealed that PBNPs did not significantly alter cell viability, proliferation, and migration activity in PBNP-labeled MSCs. Oil Red O staining and Alizarin Red staining revealed that labeled MSCs also have a normal differentiation capacity. Phalloidin staining showed no negative effect of PBNPs on the cytoskeleton. Western blot analysis indicated that PBNPs also did not change the expression of β-catenin and vimentin of MSCs. In vitro MRI, the pellets of the MSCs incubated with PBNPs showed a clear MRI signal darkening effect. In conclusion, PBNPs can be effectively used for the labeling of MSCs and will not influence the biological characteristics of MSCs