Category: Assay Development and Screening
The lack of oxygen supply is associated with a number of life-threatening diseases such as stroke, myocardial infarction or renal failure whereby cells are temporarily deprived of O2 and nutrient (ischemia). Significant cell damage can also occur during the reperfusion phase through oxidative stress and inflammatory responses. Investigating these pathologies in vitro requires an experimental set-up capable of rapid deoxygenation, rapid reperfusion, and parallel monitoring of critical biological parameters including cellular oxygenation and reactive oxygen species (ROS). The in vitro ischemia-reperfusion model presented here uses a microplate reader with software-controlled programmable O2 and CO2 regulation in combination with MitoXpress®-Intra, (Luxcel Biosciences) which enables real-time monitoring of cellular oxygenation. The system was used to study ischemia-reperfusion in HEPG2 cells and iPS derived cardiomyocytes (Cor.4U®, Axiogenesis).
The CLARIOstar microplate reader with ACU rapidly regulated the programmed decrease and increase of O2 mimicking the ischemia-reperfusion insult. Oxygenation was monitored in real-time using MitoXpress-Intra and revealed the response to changing oxygen conditions dependent on the cell’s metabolic state. In parallel, induction of reactive oxygen species and mitochondrial membrane potential was acquired. The data emphasize the suitability of the microplate reader-based model to measure multiple biological aspects in response to ischemia-reperfusion conditions.
Catherine Wark– Senior Applications Scientist, BMG LABTECH Ltd., Aylesbury, England, United Kingdom
Senior Applications Scientist
BMG LABTECH Ltd.
Aylesbury, England, United Kingdom
Catherine Wark is a Senior Applications Scientist with BMG LABTECH based in UK. She has fifteen years’ experience with BMG LABTECH, assisting users with microplate reader and assay optimization, drawing from her previous experiences in both academia and industry working in the fields of cell and molecular biology.