Isoflurane Preconditioning Elicits Competent Endogenous Mechanisms of Protection from Oxidative Stress in Cardiomyocytes Derived from Human Embryonic Stem Cells

• DOI: 10.1097/aln.0b013e3181eff6b7
• Publication Date: 2010-09-04T08:22:36Z
• Authors: Ana Sepac; Filip Sedlic; Karim Si-Tayeb; John Lough; Stephen A. Duncan; Martin Bienengraeber; Frank Park; Jinhee Kim; Zeljko J. Bosnjak
• Keywords: Microscopy, Confocal; Isoflurane; human embryonic stem cells; cardiomyocytes; preconditioning; mitochondria; experimental model; Genetic Vectors; Lentivirus; Hydrogen Peroxide; Immunohistochemistry; Mitochondria, Heart; Membrane Potentials; Oxidative Stress; 03 medical and health sciences; Oxygen Consumption; 0302 clinical medicine; KATP Channels; Anesthetics, Inhalation; Ischemic Preconditioning, Myocardial; Humans; Myocytes, Cardiac; Reactive Oxygen Species; Microdissection; Cells, Cultured; Embryonic Stem Cells

Human embryonic stem cell (hESC)-derived cardiomyocytes potentially represent a powerful experimental model complementary to myocardium obtained from patients that is relatively inaccessible for research purposes. We tested whether anesthetic-induced preconditioning (APC) with isoflurane elicits competent protective mechanisms in hESC-derived against oxidative stress be used as of human studying preconditioning.H1 hESC line was differentiated into using growth factors activin A and bone morphogenetic protein-4. Living ventricular were identified lentiviral vector expressing reporter gene (enhanced green fluorescent protein) driven by cardiac-specific myosin light chain-2v promoter. Mitochondrial membrane potential, reactive oxygen species production, opening mitochondrial permeability transition pore, survival assessed confocal microscopy. Oxygen consumption measured contracting clusters.Differentiation yielded high percentage (∼85%) beating clusters positive markers exhibited action potentials resembling those mature cardiomyocytes. Isoflurane depolarized mitochondria, attenuated consumption, stimulated generation species. APC protected these cells stress-induced death delayed pore opening.APC cardiomyocytes, suggesting the feasibility use other treatments/diseases. Our differentiation protocol very efficient yields These results also suggest promising ability protect improve engraftment ischemic heart.