The Effect of Uncoated SPIONs on hiPSC-Differentiated Endothelial Cells
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) p...
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| Published in | International journal of molecular sciences Vol. 20; no. 14; p. 3536 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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19.07.2019
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| ISSN | 1422-0067 1661-6596 1422-0067 |
| DOI | 10.3390/ijms20143536 |
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| 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. |
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| AbstractList | 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.BACKGROUNDEndothelial 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.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.METHODSECs 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.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.RESULTSWe 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.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.CONCLUSIONWe 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. 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. 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. 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. 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. 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. First preclinical studies on animal subjects report positive effects on ischemic myocardium, limb neovascularization or peripheral vascular regeneration [3,4,5]. 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. [...]uncoated nanoparticles could offer better efficiency of uptake as their size is not affected by coating. [...]while it was previously believed that uncoated SPIONs (uSPIONs) have higher toxicity compared to coated nanoparticles, later it was reported that incubation of nanoparticles in serum prevents surface reactivity and results in decreased cytotoxicity [25]. [...]we studied the effects of uSPIONs on cell characteristics desired for clinical application; specifically, their endothelial profile and angiogenic properties. 2. |
| Author | Sedlackova, Miroslava Zajickova, Lenka Koutna, Irena Matula, Pavel Veverkova, Lenka Simara, Pavel Synek, Petr Nichtova, Zuzana Salingova, Barbara Jasek, Ondrej |
| AuthorAffiliation | 6 Institute of Molecular Physiology and Genetics, Slovak Academy of Sciences, Dubravska Cesta 9, 84005 Bratislava, Slovak Republic 1 Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic 4 1st Surgical Department, Faculty of Medicine, Masaryk University and St. Ann’s Hospital, Pekarska 53, 65691 Brno, Czech Republic 3 Department of Physical Electronics, Faculty of Science, Masaryk University, Kotlarska 2, 60200 Brno, Czech Republic 5 Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic 2 RG Plasma Technologies, CEITEC—Masaryk University, Purkynova 656/123, 61200 Brno, Czech Republic |
| AuthorAffiliation_xml | – name: 5 Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic – name: 6 Institute of Molecular Physiology and Genetics, Slovak Academy of Sciences, Dubravska Cesta 9, 84005 Bratislava, Slovak Republic – name: 3 Department of Physical Electronics, Faculty of Science, Masaryk University, Kotlarska 2, 60200 Brno, Czech Republic – name: 4 1st Surgical Department, Faculty of Medicine, Masaryk University and St. Ann’s Hospital, Pekarska 53, 65691 Brno, Czech Republic – name: 2 RG Plasma Technologies, CEITEC—Masaryk University, Purkynova 656/123, 61200 Brno, Czech Republic – name: 1 Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic |
| Author_xml | – sequence: 1 givenname: Barbara surname: Salingova fullname: Salingova, Barbara – sequence: 2 givenname: Pavel orcidid: 0000-0001-7581-7048 surname: Simara fullname: Simara, Pavel – sequence: 3 givenname: Pavel surname: Matula fullname: Matula, Pavel – sequence: 4 givenname: Lenka orcidid: 0000-0002-6906-8906 surname: Zajickova fullname: Zajickova, Lenka – sequence: 5 givenname: Petr orcidid: 0000-0002-9590-2562 surname: Synek fullname: Synek, Petr – sequence: 6 givenname: Ondrej surname: Jasek fullname: Jasek, Ondrej – sequence: 7 givenname: Lenka surname: Veverkova fullname: Veverkova, Lenka – sequence: 8 givenname: Miroslava surname: Sedlackova fullname: Sedlackova, Miroslava – sequence: 9 givenname: Zuzana surname: Nichtova fullname: Nichtova, Zuzana – sequence: 10 givenname: Irena orcidid: 0000-0002-1680-5052 surname: Koutna fullname: Koutna, Irena |
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| CitedBy_id | crossref_primary_10_3390_nano12091414 crossref_primary_10_1093_mam_ozae048 crossref_primary_10_4103_jpbs_jpbs_846_24 crossref_primary_10_3390_pharmaceutics16091109 crossref_primary_10_1016_j_isci_2022_104947 |
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| Keywords | reprogramming human induced pluripotent stem cell-derived endothelial cells differentiation superparamagnetic iron-oxide nanoparticles mature endothelial cells |
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| SubjectTerms | Apoptosis Biomarkers Cell culture Cell Differentiation Cell Survival Cells, Cultured Cytotoxicity Endothelial Cells - cytology Endothelial Cells - metabolism Endothelial Cells - ultrastructure Ferric Compounds - chemistry Human Umbilical Vein Endothelial Cells Humans Immunohistochemistry Induced Pluripotent Stem Cells - cytology Induced Pluripotent Stem Cells - metabolism Induced Pluripotent Stem Cells - ultrastructure Labeling Magnetite Nanoparticles - chemistry Metabolism Nanoparticles Population Transmission electron microscopy |
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| Title | The Effect of Uncoated SPIONs on hiPSC-Differentiated Endothelial Cells |
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