BACKGROUND: Endothelial progenitor cells (EPCs) were indicated in vascular repair, angiogenesis of ischemic organs, and inhibition of formation of initial hyperplasia. Differentiation of endothelial cells (ECs) from human induced pluripotent stem cells (hiPSC)-derived endothelial cells (hiPSC-ECs) provides an unlimited supply for clinical application. Furthermore, magnetic cell labelling offers an effective way of targeting and visualization of hiPSC-ECs and is the next step towards in vivo studies. METHODS: ECs were differentiated from hiPSCs and labelled with uncoated superparamagnetic iron-oxide nanoparticles (uSPIONs). uSPION uptake was compared between hiPSC-ECs and mature ECs isolated from patients by software analysis of microscopy pictures after Prussian blue cell staining. The acute and long-term cytotoxic effects of uSPIONs were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay) and Annexin assay. RESULTS: We showed, for the first time, uptake of uncoated SPIONs (uSPIONs) by hiPSC-ECs. In comparison with mature ECs of identical genetic background hiPSC-ECs showed lower uSPION uptake. However, all the studied endothelial cells were effectively labelled and showed magnetic properties even with low labelling concentration of uSPIONs. uSPIONs prepared by microwave plasma synthesis did not show any cytotoxicity nor impair endothelial properties. CONCLUSION: We show that hiPSC-ECs labelling with low concentration of uSPIONs is feasible and does not show any toxic effects in vitro, which is an important step towards animal studies.

1st Surgical Department Faculty of Medicine Masaryk University and St Ann's Hospital Pekarska 53 65691 Brno Czech Republic

Centre for Biomedical Image Analysis Faculty of Informatics Masaryk University Kamenice 5 62500 Brno Czech Republic

Department of Histology and Embryology Faculty of Medicine Masaryk University Kamenice 5 62500 Brno Czech Republic

Department of Physical Electronics Faculty of Science Masaryk University Kotlarska 2 60200 Brno Czech Republic

Institute of Molecular Physiology and Genetics Slovak Academy of Sciences Dubravska Cesta 9 84005 Bratislava Slovak Republic

RG Plasma Technologies CEITEC Masaryk University Purkynova 656 123 61200 Brno Czech Republic

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