An in vitro model for assessment of recovery of cardiomyocytes from hypoxia/reperfusion injury

Reperfusion of cardiac tissue is necessary to resuscitate ischemic or hypoxic myocardium after myocardial infarction (MI). Restoration, however, often results in a number of deleterious effects termed reperfusion injury. This work presents an in vitro model for assessing the recovery of fetal canine cardiomyocytes from hypoxia/reperfusion injury.;The in vitro experimental model introduced in this study enables the side-by-side culture of normal and compromised, myocytes in the same monolayer. This model consists of creating a region of hypoxia by flowing anoxic media over a portion of the culture and oxygenated media over the remainder. Low-Reynolds-number fluid dynamic manipulation is used to provide control of the cell culture environment. Injured myocytes undergo significant morphological changes upon reoxygenation. Fluorescence microscopy reveals elevated levels of apoptosis (∼ 20%) attributed to reperfusion injury. In contrast with existing models of hypoxia/ischemia, this model provides a simple and effective way to create hypoxic/ischemic conditions in vitro, to study changes along side of a control within the same monolayer, and to access different therapeutic treatments (cells, media supplements, etc.) on injured and healthy cells.;Cell-based therapies are a distinct class of treatments that rely on cells as the therapeutic agents to affect a pathological condition. Injection of stem and stromal cells into injured myocardium have revealed significant improvement in animal and human models, however, the underlying mechanisms have not been identified. To this end, conditioned media was collected after two days of incubation with adipose-derived stromal cells under normal conditions and introduced into the bioreactor during reoxygenation. No significant changes in apoptosis or necrosis occurred due to the addition of conditioned media at the concentrations used in the investigation.