EdU sensing: The Raman way of following endothelial cell proliferation in vitro and ex vivo

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dc.contributor.authorRadwan, Basseem
dc.contributor.authorRocchetti, Stefano
dc.contributor.authorMatuszyk, Ewelina
dc.contributor.authorSternak, Magdalena
dc.contributor.authorStodulski, Maciej
dc.contributor.authorPawlowski, Robert
dc.contributor.authorMlynarski, Jacek
dc.contributor.authorBrzozowski, Krzysztof
dc.contributor.authorChlopicki, Stefan
dc.contributor.authorBaranska, Malgorzata
dc.contributor.organizationJagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian Universityen
dc.contributor.organizationFaculty of Chemistry, Jagiellonian Universityen
dc.contributor.organizationInstitute of Organic Chemistry, Polish Academy of Sciencesen
dc.date.accessioned2022-09-14T12:25:30Z
dc.date.available2022-09-14T12:25:30Z
dc.date.issued2022
dc.description.abstractEndothelial cells line the lumen of all vessels in the body and maintain vascular homeostasis. In particular, endothelial cell regeneration in response to insult sustain functional endothelial layer. EdU (5-ethynyl-2′-deoxyuridine) is an alkyne-tagged proliferation probe that incorporates into newly synthesized DNA and is used for fluorescence imaging of cell proliferation with the use of “click chemistry” reaction with a fluorescent azide. Here, we utilized EdU as a click-free Raman probe for tracking endothelial cell proliferation. Raman imaging of EdU was performed in live endothelial cells, showing an advantage over fluorescence imaging of EdU, as this technique did not require sample fixation and permeabilization. To validate Raman-based imaging of EdU to study endothelial cell proliferation, we showed that when endothelial cells were treated with cycloheximide or doxorubicin to impair the proliferation of endothelial cells, the Raman-based signal of EdU was diminished. Furthermore, endothelial cells proliferation detected using EdU-labelled Raman imaging was compared with fluorescence imaging. Finally, the method of Raman–based EdU imaging was used in the isolated murine aorta ex vivo. Altogether, our results show that Raman-based imaging of EdU provides a novel alternative for fluorescence-based assay to assess endothelial proliferation and regeneration.en
dc.description.sponsorshipEuropean Union's Horizon 2020 Research and Innovation Programme under Marie Sklodowska-Curie [Grant Agreement 813920 (LogicLab ITN)] National Science Center Poland (NCN) [OPUS15 no. UMO-2018/29/B/ST4/00335 to MB]
dc.identifier.citationBiosensors and Bioelectronics Volume 216, 15 November 2022, 114624 ; https://doi.org/10.1016/j.bios.2022.114624en
dc.identifier.doi10.1016/j.bios.2022.114624
dc.identifier.issn0956-5663
dc.identifier.urihttps://open.icm.edu.pl/handle/123456789/21629
dc.language.isoen
dc.publisherElsevieren
dc.rightsUznanie autorstwa 4.0 Międzynarodowe*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectRaman imagingen
dc.subjectclick chemistryen
dc.subjectfluorescence imagingen
dc.subjectendotheliumen
dc.subjectcell proliferationen
dc.subjectcycloheximideen
dc.subjectdoxorubicinen
dc.titleEdU sensing: The Raman way of following endothelial cell proliferation in vitro and ex vivoen
dc.typearticleen
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