Projects per year
Abstract
Reactive oxygen species (ROS) have different properties and biological functions. They contribute to cell signaling and, in excessive amounts, to oxidative stress (OS). Although ROS is pivotal in a wide number of physiological systems and pathophysiological processes, direct quantification in vivo is quite challenging and mainly limited to in vitro studies. Even though advanced in vitro cell culture techniques, like on-a-chip culture, have overcome the lack of crucial in vivo-like physiological aspects in 2D culture, the majority of in vitro ROS quantification studies are generally performed in 2D. Here we report the development, application, and validation of a multiplexed assay to quantify ROS and cell viability in organ-on-a-chip models. The assay utilizes three dyes to stain live cells for ROS, dead cells, and DNA. Confocal images were analyzed to quantify ROS probes and determine the number of nuclei and dead cells. We found that, in contrast to what has been reported with 2D cell culture, on-a-chip models are more prone to scavenge ROS rather than accumulate them. The assay is sensitive enough to distinguish between different phenotypes of endothelial cells (ECs) based on the level of OS to detect higher level in tumor than normal cells. Our results indicate that the use of physiologically relevant models and this assay could help unravelling the mechanisms behind OS and ROS accumulation. A further step could be taken in data analysis by implementing AI in the pipeline to also analyze images for morphological changes to have an even broader view of OS mechanism.
Original language | English |
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Article number | 102488 |
Number of pages | 10 |
Journal | Redox Biology |
Volume | 57 |
Early online date | 29 Sept 2022 |
DOIs | |
Publication status | Published - Nov 2022 |
Keywords
- Angiogenesis
- Antioxidant
- Cell viability
- Multiplex assay
- Organ-on-a-chip
- Oxidative stress
- ROS
ASJC Scopus subject areas
- Organic Chemistry
- Clinical Biochemistry
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Dive into the research topics of 'A versatile multiplexed assay to quantify intracellular ROS and cell viability in 3D on-a-chip models'. Together they form a unique fingerprint.Projects
- 1 Finished
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Innovation in modelling Placenta for Maternal and Fetal Health (iPLACENTA) (joint with Aston University, Mimetas BV, St George's Medical School, University College Cork, Universita Degli Studi di Torino, Institut National de le Sante et de la Recherche Medicale, Universitaet Rostock, Fundacion Para le Investigacion del Hospital Universitario La Fe De La Comunidad Valencia, Katholieke Universiteit Leuven, Universiteit Maastricht)
Murdoch, C. (Investigator)
COMMISSION OF THE EUROPEAN COMMUNITIES
1/03/18 → 31/05/22
Project: Research
Activities
- 1 Funding - grants and income which support research related activities
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Innovation in modelling Placenta for Maternal and Fetal Health (iPLACENTA) (joint with Mimetas BV, St George's Medical School, University College Cork, Universita Degli Studi di Torino, Institut National de le Sante et de la Recherche Medicale, Universitaet Rostock, Fundacion Para le Investigacion del Hospital Universitario La Fe De La Comunidad Valencia, Aston University, Katholieke Universiteit Leuven, Universiteit Maastricht)
Murdoch, C. (Recipient)
1 Mar 2018 → 31 May 2022Activity: Other activity types › Funding - grants and income which support research related activities