Abstract
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.
Highlights
Endothelial cell therapies show great potential in treatment of multiple diseases
endothelial cells (ECs) were differentiated from human induced pluripotent stem cells (hiPSC) and labelled with uncoated superparamagnetic iron-oxide nanoparticles. uncoated SPIONs (uSPIONs) uptake was compared between hiPSC-ECs and mature ECs isolated from patients by software analysis of microscopy pictures after Prussian blue cell staining
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
Summary
Endothelial cell therapies show great potential in treatment of multiple diseases. Endothelial progenitor cells (EPCs) were indicated in vascular repair, angiogenesis of ischemic organs, and inhibiting the formation of intimal hyperplasia [1,2]. Cell labelling is a convenient tool for detailed study of endothelial cell function in vivo, and cell tracking and guiding mechanisms for regeneration therapy. It can provide valuable information about membrane properties of hiPSC-ECs, which were not studied until now. In some cases an iron-oxide core of size around 10 nm can result in a nanoparticles ranging from 80 nm to 150 nm dependent on organic coating, which significantly affects the efficiency of uptake and magnetic properties of the labelled cell [22,23,24]. We studied the effects of uSPIONs on cell characteristics desired for clinical application; their endothelial profile and angiogenic properties
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