Abstract
The paper presents a newly designed microfluidic system that allows simulation of myocardial hypoxia by biochemical method. The geometry of the microsystem was designed in such a way, that quantitative fluorescent measurements using a spectrofluorometric plate reader was possible. Biochemical simulation of hypoxia was carried out using potent mitochondrial oxidative phosphorylation uncoupler—Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP). Two cardiac cell lines were used in the study—rat cardiomyoblasts (H9C2) and human cardiomyocytes. The effectiveness of biochemical simulation of hypoxia was studied using two fluorescent dyes: carbocyanine iodide (JC-1) and Fluo-4. Changes in the mitochondrial membrane potential and concentration of intracellular calcium ions were tested. The major novelty of this research was the applying the microfluidic system to create hypoxia conditions for cardiac cells using the biochemical approach. In further studies, the presented hypoxia model could be used to develop new methods of treatment of ischemic heart disease for example in cell therapy based on stem cells.
Highlights
The paper presents a newly designed microfluidic system that allows simulation of myocardial hypoxia by biochemical method
Microfluidic systems could be useful tools in the development of the cellular model of Ischemic heart disease (IHD). These microsystems have a lot of advantages such as: the possibility of designing geometry and dimensions mimicking the natural microenvironment of the cells, control of cell culture conditions, physical parameters similar to those occurring in physiological conditions (surface-to-volume ratio (SAV), an effective culture volume (ECV), laminar flow), the possibility of conducting real-time analysis and they are an alternative tool to animal studies[5]
Optimization of the stimulation of myocardial ischemia on two cardiac cell lines was performed in the macroscale
Summary
The paper presents a newly designed microfluidic system that allows simulation of myocardial hypoxia by biochemical method. Researchers have tended to develop models of diseases, including IHD For this purpose, it is necessary to create hypoxic conditions in the microsystems. At the time of removing the cells from the hypoxia chamber for testing, oxygen from the atmosphere begins to diffuse into the medium, which changes the culture conditions. Another widely used solution to simulate hypoxia in a microsystem are the gas channels. They might have a negative effect on cell viability, metabolism or morphology
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