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Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform
Cellular activation and inflammation leading to endothelial dysfunction is associated with cardiovascular disease (CVD). We investigated whether a single cell label-free multi parameter optical interrogation system can detect endothelial cell and endothelial progenitor cell (EPC) activation in vitro...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411894/ https://www.ncbi.nlm.nih.gov/pubmed/30858536 http://dx.doi.org/10.1038/s41598-019-40612-8 |
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author | King, Damien Glynn, MacDara Cindric, Sandra Kernan, David O’Connell, Tríona Hakimjavadi, Roya Kearney, Sinéad Ackermann, Tobias Berbel, Xavier Munoz Llobera, Andreu Simonsen, Ulf Laursen, Britt E. Redmond, Eileen M. Cahill, Paul A. Ducrée, Jens |
author_facet | King, Damien Glynn, MacDara Cindric, Sandra Kernan, David O’Connell, Tríona Hakimjavadi, Roya Kearney, Sinéad Ackermann, Tobias Berbel, Xavier Munoz Llobera, Andreu Simonsen, Ulf Laursen, Britt E. Redmond, Eileen M. Cahill, Paul A. Ducrée, Jens |
author_sort | King, Damien |
collection | PubMed |
description | Cellular activation and inflammation leading to endothelial dysfunction is associated with cardiovascular disease (CVD). We investigated whether a single cell label-free multi parameter optical interrogation system can detect endothelial cell and endothelial progenitor cell (EPC) activation in vitro and ex vivo, respectively. Cultured human endothelial cells were exposed to increasing concentrations of tumour necrosis factor alpha (TNF-α) or lipopolysaccharide (LPS) before endothelial activation was validated using fluorescence-activated cell sorting (FACS) analysis of inflammatory marker expression (PECAM-1, E-selectin and ICAM-1). A centrifugal microfluidic system and V-cup array was used to capture individual cells before optical measurement of light scattering, immunocytofluorescence, auto-fluorescence (AF) and cell morphology was determined. In vitro, TNF-α promoted specific changes to the refractive index and cell morphology of individual cells concomitant with enhanced photon activity of fluorescently labelled inflammatory markers and increased auto-fluorescence (AF) intensity at three different wavelengths, an effect blocked by inhibition of downstream signalling with Iκβ. Ex vivo, there was a significant increase in EPC number and AF intensity of individual EPCs from CVD patients concomitant with enhanced PECAM-1 expression when compared to normal controls. This novel label-free ‘lab on a disc’ (LoaD) platform can successfully detect endothelial activation in response to inflammatory stimuli in vitro and ex vivo. |
format | Online Article Text |
id | pubmed-6411894 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-64118942019-03-13 Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform King, Damien Glynn, MacDara Cindric, Sandra Kernan, David O’Connell, Tríona Hakimjavadi, Roya Kearney, Sinéad Ackermann, Tobias Berbel, Xavier Munoz Llobera, Andreu Simonsen, Ulf Laursen, Britt E. Redmond, Eileen M. Cahill, Paul A. Ducrée, Jens Sci Rep Article Cellular activation and inflammation leading to endothelial dysfunction is associated with cardiovascular disease (CVD). We investigated whether a single cell label-free multi parameter optical interrogation system can detect endothelial cell and endothelial progenitor cell (EPC) activation in vitro and ex vivo, respectively. Cultured human endothelial cells were exposed to increasing concentrations of tumour necrosis factor alpha (TNF-α) or lipopolysaccharide (LPS) before endothelial activation was validated using fluorescence-activated cell sorting (FACS) analysis of inflammatory marker expression (PECAM-1, E-selectin and ICAM-1). A centrifugal microfluidic system and V-cup array was used to capture individual cells before optical measurement of light scattering, immunocytofluorescence, auto-fluorescence (AF) and cell morphology was determined. In vitro, TNF-α promoted specific changes to the refractive index and cell morphology of individual cells concomitant with enhanced photon activity of fluorescently labelled inflammatory markers and increased auto-fluorescence (AF) intensity at three different wavelengths, an effect blocked by inhibition of downstream signalling with Iκβ. Ex vivo, there was a significant increase in EPC number and AF intensity of individual EPCs from CVD patients concomitant with enhanced PECAM-1 expression when compared to normal controls. This novel label-free ‘lab on a disc’ (LoaD) platform can successfully detect endothelial activation in response to inflammatory stimuli in vitro and ex vivo. Nature Publishing Group UK 2019-03-11 /pmc/articles/PMC6411894/ /pubmed/30858536 http://dx.doi.org/10.1038/s41598-019-40612-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article King, Damien Glynn, MacDara Cindric, Sandra Kernan, David O’Connell, Tríona Hakimjavadi, Roya Kearney, Sinéad Ackermann, Tobias Berbel, Xavier Munoz Llobera, Andreu Simonsen, Ulf Laursen, Britt E. Redmond, Eileen M. Cahill, Paul A. Ducrée, Jens Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title | Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title_full | Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title_fullStr | Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title_full_unstemmed | Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title_short | Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in Single Cells using a Lab on a Disc Platform |
title_sort | label-free multi parameter optical interrogation of endothelial activation in single cells using a lab on a disc platform |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6411894/ https://www.ncbi.nlm.nih.gov/pubmed/30858536 http://dx.doi.org/10.1038/s41598-019-40612-8 |
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