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First published online January 25, 2007
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2006-0466v1
25/5/1136    most recent
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Submitted on July 25, 2006
Accepted on January 11, 2007

Embryonic Stem Cells

Developmental changes in cardiomyocytes differentiated from human embryonic stem cells: a molecular and electrophysiological approach

Laura Sartiani 1, Esther Bettiol 2, Francesca Stillitano 1, Alessandro Mugelli 1, Elisabetta Cerbai 1, Marisa E. Jaconi 2*

1 Centro Interuniversitario di Medicina Molecolare e Biofisica Applicata (C.I.M.M.B.A.), University of Firenze, Firenze, Italy
2 Biology of Aging Laboratory, Department of Rehabilitation and Geriatrics, University Hospital of Geneva, Geneva, Switzerland

* To whom correspondence should be addressed. E-mail: Marisa.Jaconi{at}medecine.unige.ch.


  Abstract

Cardiomyocytes derived from human embryonic stem cells constitute a promising cell source for the regeneration of damaged hearts. The assessment of their in vitro functional properties is mandatory to envisage appropriate cardiac cell-based therapies. In this study, we characterized human embryonic stem cell-derived cardiomyocytes over a three-month period, using patch-clamp or intracellular recordings to assess their functional maturation and RT-PCR to evaluate the expression of ion channel-encoding subunits. Ito1 and IK1, the transient outward and inward rectifier potassium currents, were present in cardiomyocytes only, whereas IKr, the rapid delayed rectifier potassium current, If, the pacemaker current and ICa,L, the L-type calcium current, could be recorded both in undifferentiated human embryonic stem cells and cardiomyocytes. Most of the currents underwent developmental maturation in cardiomyocytes, as assessed by modifications in current density (Ito1, IK1 and ICa,L) and properties (If). Ion channel mRNAs were always present when the current was recorded. Intracellular recordings in spontaneously beating clusters of cardiomyocytes revealed changes in action potential parameters and in response to pharmacological tools according to time of differentiation. In summary, human embryonic stem cell-derived cardiomyocytes mature over time during in vitro differentiation, approaching an adult phenotype.

Key Words. ion current, maturation, ion channel subunits, multicellular recordings




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