Isoflurane preconditioning elicits competent endogenous mechanisms of protection from oxidative stress in cardiomyocytes derived from human embryonic stem cells

Ana Sepac, Filip Sedlic, Karim Si-Tayeb, John Lough, Stephen A. Duncan, Martin Bienengraeber, Frank Park, Jinhee Kim, Zeljko J. Bosnjak

Research output: Contribution to journalArticle

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Abstract

Background: 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 cardiomyocytes against oxidative stress to be used as a model of human cardiomyocytes for studying preconditioning. Methods: H1 hESC cell line was differentiated into cardiomyocytes using growth factors activin A and bone morphogenetic protein-4. Living ventricular hESC-derived cardiomyocytes were identified using a lentiviral vector expressing a reporter gene (enhanced green fluorescent protein) driven by a cardiac-specific human myosin light chain-2v promoter. Mitochondrial membrane potential, reactive oxygen species production, opening of mitochondrial permeability transition pore, and survival of hESC-derived cardiomyocytes were assessed using confocal microscopy. Oxygen consumption was measured in contracting cell clusters. Results: Differentiation yielded a high percentage (β85%) of cardiomyocytes in beating clusters that were positive for cardiac-specific markers and exhibited action potentials resembling those of mature cardiomyocytes. Isoflurane depolarized mitochondria, attenuated oxygen consumption, and stimulated generation of reactive oxygen species. APC protected these cells from oxidative stress-induced death and delayed mitochondrial permeability transition pore opening. Conclusions: APC elicits competent protective mechanisms against oxidative stress in hESC-derived cardiomyocytes, suggesting the feasibility to use these cells as a model of human cardiomyocytes for studying APC and potentially other treatments/diseases. Our differentiation protocol is very efficient and yields a high percentage of cardiomyocytes. These results also suggest a promising ability of APC to protect and improve engraftment of hESC-derived cardiomyocytes into the ischemic heart.

Original languageEnglish (US)
Pages (from-to)906-916
Number of pages11
JournalAnesthesiology
Volume113
Issue number4
DOIs
StatePublished - Jan 1 2010

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Isoflurane
Cardiac Myocytes
Oxidative Stress
Anesthetics
Oxygen Consumption
Human Embryonic Stem Cells
Reactive Oxygen Species
Bone Morphogenetic Protein 4
Myosin Light Chains
Mitochondrial Membrane Potential
Reporter Genes
Confocal Microscopy
Action Potentials
Intercellular Signaling Peptides and Proteins
Myocardium
Mitochondria
Theoretical Models

All Science Journal Classification (ASJC) codes

  • Anesthesiology and Pain Medicine

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Isoflurane preconditioning elicits competent endogenous mechanisms of protection from oxidative stress in cardiomyocytes derived from human embryonic stem cells. / Sepac, Ana; Sedlic, Filip; Si-Tayeb, Karim; Lough, John; Duncan, Stephen A.; Bienengraeber, Martin; Park, Frank; Kim, Jinhee; Bosnjak, Zeljko J.

In: Anesthesiology, Vol. 113, No. 4, 01.01.2010, p. 906-916.

Research output: Contribution to journalArticle

Sepac, Ana ; Sedlic, Filip ; Si-Tayeb, Karim ; Lough, John ; Duncan, Stephen A. ; Bienengraeber, Martin ; Park, Frank ; Kim, Jinhee ; Bosnjak, Zeljko J. / Isoflurane preconditioning elicits competent endogenous mechanisms of protection from oxidative stress in cardiomyocytes derived from human embryonic stem cells. In: Anesthesiology. 2010 ; Vol. 113, No. 4. pp. 906-916.
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AU - Sedlic, Filip

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AU - Lough, John

AU - Duncan, Stephen A.

AU - Bienengraeber, Martin

AU - Park, Frank

AU - Kim, Jinhee

AU - Bosnjak, Zeljko J.

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