Copyright © 2006, European Society of Cardiology
Studying ischemic preconditioning in isolated cardiomyocyte models
aDivision of Cardiovascular Research, McMaster Building, Room 7019C, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada M4B 1E1
bDepartment of Laboratory Medicine and Pathobiology, The Hospital for Sick Children and The University of Toronto, Toronto, Canada
* Corresponding author. Tel.: +1 416 813 8720; fax: +1 416 813 7480. Email address: Diazport{at}sickkids.ca
Isolated cardiomyocytes, obtained by enzymatic digestion of whole hearts, have multiple advantages, most related to their accessibility to microscopic visualization, beyond the obvious elimination of other cell types that exist in the heart. Conversely, they cannot reproduce the mechanical disruption of reperfusion hypercontracture or the vascular phenomena of leukocyte plugging and compression from interstitial edema and contracture that can lead to the no-reflow phenomenon. Nevertheless, ischemic preconditioning has been consistently demonstrated to be a potent protective mechanism in freshly isolated and cultured cardiomyocytes across multiple species, indicating that much of the innate protection of ischemic preconditioning resides in cardiomyocytes. Centrifuging freshly isolated cardiomyocytes into a pellet with only a thin layer of supernatant covered by oil has proven to be an excellent model of simulated ischemia. In culture, cardiomyocytes may be exposed to severe hypoxia only or to various protocols for simulated ischemia in which an acid/lactate-rich, hyperkalemic extracellular environment with substrate deprivation (lacking glucose) is typically added. Reperfusion is simulated by well-oxygenated media of normal ionic composition. Cardiomyocyte injury has been usually evaluated by cell membrane permeability to dyes, often under hypo-osmotic conditions (osmotic fragility) or enzyme release. A survey of the use of cardiomyocyte models to study preconditioning is presented with the emphasis on examples of the innovative measurements, increasingly involving molecular techniques, that point to an increasing future role for these models in preconditioning research and, more generally, in the mechanistic study of myocardial ischemia/reperfusion.
KEYWORDS Ischemic preconditioning; Simulated ischemia/reperfusion; Cardiomyocytes; Cell culture
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