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1 Department of Physiology, Loyola University Chicago, Maywood, IL, USA; Institut fuer Neurophysiologie, Universitaet zu Koeln, Koeln, Germany
2 Institut fuer Neurophysiologie, Universitaet zu Koeln, Koeln, Germany
3 Neurobiologie und Biophysik, Institut fuer Biologie III, Albert-Ludwigs Universitaet Freiburg, Freiburg, Germany
* To whom correspondence should be addressed. E-mail: kbanac1{at}lumc.edu.
Embryonic stem cells differentiate into cardiac myocytes, repeating in-vitro the structural and molecular changes associated with cardiac development. Currently it is not clear whether the electrophysiological properties of the multicellular cardiac structure follow cardiac maturation as well. In long-term recordings of extracellular field potentials with microelectrode arrays (MEA) consisting of 60 substrate-integrated electrodes, we examined the electrophysiological properties during the ongoing differentiation process. The beating frequency of the growing preparations increased from 1 to 5Hz concomitant to a decrease of the AP duration and the AP rise time. A developmental increase of the conduction velocity could be attributed to an increased expression of connexin43 (Cx43) gap junction channels. Whereas isoprenalin elicited a positive chronotropic response from the first day of spontaneous beating onward, a concentration dependent negative chronotropic effect of carbachol only developed after ~4 days. The in-vitro development of the 3D cardiac preparation thus closely follows the development described for mouse embryonic heart, making it an ideal model to monitor the differentiation of electrical activity in embryonic cardiomyocytes.
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