Universität zu Köln

Hannes, Tobias ORCID: 0000-0002-8550-2282, Wolff, Marie, Doss, Michael Xavier, Pfannkuche, Kurt, Haustein, Moritz, Mueller-Ehmsen, Jochen, Sachinidis, Agapios, Hescheler, Juergen, Khalil, Markus and Halbach, Marcel (2015). Electrophysiological Characteristics of Embryonic Stem Cell-Derived Cardiomyocytes are Cell Line-Dependent. Cell. Physiol. Biochem., 35 (1). S. 305 - 315. BASEL: KARGER. ISSN 1421-9778

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Abstract

Background: Modelling of cardiac development, physiology and pharmacology by differentiation of embryonic stem cells (ESCs) requires comparability of cardiac differentiation between different ESC lines. To investigate whether the outcome of cardiac differentiation is consistent between different ESC lines, we compared electrophysiological properties of ESC-derived cardiomyocytes (ESC-CMs) of different murine ESC lines. Methods: Two wild-type (D3 and R1) and two transgenic ESC lines (D3/aPIG44 and CGR8/AMPIGX-7) were differentiated under identical culture conditions. The transgenic cell lines expressed enhanced green fluorescent protein (eGFP) and puromycin-N-acetyltransferase under control of the cardiac specific alpha-myosin heavy chain (alpha MHC) promoter. Action potentials (APs) were recorded using sharp electrodes and multielectrode arrays in beating clusters of ESC-CMs. Results: Spontaneous AP frequency and AP duration (APD) as well as maximal upstroke velocity differed markedly between unpurified CMs of the four ESC lines. APD heterogeneity was negligible in D3/aPIG44, moderate in D3 and R1 and extensive in CGR8/AMPIGX-7. Interspike intervals calculated from long-term recordings showed a high degree of variability within and between recordings in CGR8/AMPIGX-7, but not in D3/aPIG44. Purification of the aMHC+ population by puromycin treatment posed only minor changes to APD in D3/aPIG44, but significantly shortened APD in CGR8/AMPIGX-7. Conclusion: Electrophysiological properties of ESC-CMs are strongly cell line-dependent and can be influenced by purification of cardiomyocytes by antibiotic selection. Thus, conclusions on cardiac development, physiology and pharmacology derived from single stem cell lines have to be interpreted carefully. Copyright (C) 2015 S. Karger AG, Basel

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Hannes, Tobias UNSPECIFIED orcid.org/0000-0002-8550-2282 UNSPECIFIED Wolff, Marie UNSPECIFIED UNSPECIFIED UNSPECIFIED Doss, Michael Xavier UNSPECIFIED UNSPECIFIED UNSPECIFIED Pfannkuche, Kurt UNSPECIFIED UNSPECIFIED UNSPECIFIED Haustein, Moritz UNSPECIFIED UNSPECIFIED UNSPECIFIED Mueller-Ehmsen, Jochen UNSPECIFIED UNSPECIFIED UNSPECIFIED Sachinidis, Agapios UNSPECIFIED UNSPECIFIED UNSPECIFIED Hescheler, Juergen UNSPECIFIED UNSPECIFIED UNSPECIFIED Khalil, Markus UNSPECIFIED UNSPECIFIED UNSPECIFIED Halbach, Marcel UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-417607
DOI:	10.1159/000369697
Journal or Publication Title:	Cell. Physiol. Biochem.
Volume:	35
Number:	1
Page Range:	S. 305 - 315
Date:	2015
Publisher:	KARGER
Place of Publication:	BASEL
ISSN:	1421-9778
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language IN-VITRO; DIFFERENTIATION; ESTABLISHMENT; ENGRAFTMENT; EXPRESSION; CHANNELS; CULTURE Multiple languages Cell Biology; Physiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/41760