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Beta(1)- and beta(2)-adrenoceptor responses in cardiomyocytes derived from human embryonic stem cells: comparison with failing and non-failing adult human heart.

ABSTRACT: Characterization of human embryonic stem cell-derived cardiomyocytes (hESC-CM) in relation to adult myocytes is essential for their future use in transplantation or as a model system. The beta-adrenoceptor pathways, which are known to be effective early in hESC-CM development, are of major importance because of their control of rate and force of beating, arrhythmia generation and apoptosis/necrosis. We have therefore performed detailed pharmacological analysis of the beta-adrenoceptor responses in developing hESC-CM.hESC-CMs were differentiated from H7 ESCs and studied up to 79 days of differentiation. Rate of beating and time course of contraction and relaxation were measured in superfused preparations.Responses to the mixed beta(1)- and beta(2)-adrenoceptor agonist isoprenaline were evident from day 10 to day 79. Stability of the responses during an application, for repeated applications on the same experimental day and over the time of development, was determined. Concentrations for half-maximal response (12.9 nM) were similar to those from adult human heart, but closer to those obtained from failing rather than normal ventricle. Acceleration of both contraction and relaxation was quantitatively similar to that in adult ventricular myocytes, as was sensitivity to muscarinic inhibition. Use of specific antagonists showed that both beta(1)- and beta(2)-adrenoceptors contributed to contractile responses, as seen with adult myocytes.These data show the compatibility of hESC-CM with adult human myocardium in terms of beta-adrenoceptor response. The experiments described here also confirm the utility of the hESC-CM preparation for detailed pharmacological analysis.

SUBMITTER: Brito-Martins M

PROVIDER: S-EPMC2259205 | biostudies-other | 2008 Feb

REPOSITORIES: biostudies-other

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Project description:Various types of cardiomyocytes undergo changes in automaticity and electrical properties during fetal heart development. Human embryonic stem cell-derived cardiomyocytes (hESC-CMs), like fetal cardiomyocytes, are electrophysiologically immature and exhibit automaticity. We used hESC-CMs to investigate developmental changes in mechanisms of automaticity and to determine whether electrophysiological maturation is driven by an intrinsic developmental clock and/or is regulated by interactions with non-cardiomyocytes in embryoid bodies (EBs). We isolated pure populations of hESC-CMs from EBs by lentivirus-engineered Puromycin resistance at various stages of differentiation. Using pharmacological agents, calcium (Ca(2+)) imaging, and intracellular recording techniques, we found that intracellular Ca(2+)-cycling mechanisms developed early and contributed to dominant automaticity throughout hESC-CM differentiation. Sarcolemmal ion channels evolved later upon further differentiation within EBs and played an increasing role in controlling automaticity and electrophysiological properties of hESC-CMs. In contrast to the development of intracellular Ca(2+)-handling proteins, ion channel development and electrophysiological maturation of hESC-CMs did not occur when hESC-CMs were isolated from EBs early and maintained in culture without further interaction with non-cardiomyocytes. Adding back non-cardiomyocytes to early-isolated hESC-CMs rescued the arrest of electrophysiological maturation, indicating that non-cardiomyocytes in EBs drive electrophysiological maturation of early hESC-CMs. Non-cardiomyocytes in EBs contain most cell types present in the embryonic heart that are known to influence early cardiac development. Our study is the first to demonstrate that non-cardiomyocytes influence electrophysiological maturation of early hESC-CMs in cultures. Defining the nature of these extrinsic signals will aid in the directed maturation of immature hESC-CMs to mitigate arrhythmogenic risks of cell-based therapies.

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Beta(1)- and beta(2)-adrenoceptor responses in cardiomyocytes derived from human embryonic stem cells: comparison with failing and non-failing adult human heart.

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