Tissue and Cell 45 (2013) 101–106 Contents lists available at SciVerse ScienceDirect Tissue and Cell jou rn al h om epage: www.elsevier.com/locate/tice The influence of dexamethasone administration on the protection against doxorubicin-induced cardiotoxicity in purified embryonic stem cell-derived cardiomyocytes Leila Dehghani a,b , Mahbobeh Farokhpour a , Khadije Karbalaie a , Marzie Nematollahi a , Somayeh Tanhaie a , Nasim Hayati-Rodbari b , Abbas Kiani-Esfahani a , Juergen Hescheler c , Mohammad Hossein Nasr-Esfahani a,∗ , Hossein Baharvand d,e,∗∗ a Department of Cell and Molecular Biology, Cell Science Research Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran b Department of Biology, Islamic Azad University (IAU), Science and Research Branch, Tehran, Iran c Institute of Neurophysiology, University of Koln, Robert-Kock St., Koln, 39D-50931 Koln, Germany d Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran e Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran a r t i c l e i n f o Article history: Received 13 July 2012 Accepted 27 September 2012 Available online 9 November 2012 Keywords: Embryonic stem cells Cardiomyocyte differentiation Cytotoxicity Doxorubicin Dexamethasone Mifepristone a b s t r a c t Embryonic stem cells (ESCs) have various uses in drug toxicity, as they can be easily differentiated in vitro. However, one of the major obstacles in the assessment of these differentiated cells is the presence of a heterogeneous cell population. To circumvent this problem, purified ESC-derived desired cells by means of the tissue-specific GFP and/or antibiotic resistance gene expression has been proposed. Therefore, this study aimed to assess the role of doxorubicin (DOX) in cardiotoxicity by using genetically engineered purified ESC-derived cardiomyocytes under the control alpha-myosin heavy chain promoter. The results revealed that ESCs are suitable for evaluation of DOX cardiotoxicity. This study showed that DOX car- diotoxicity was reduced as detected by beating cardiomyocytes and caspase activity only by pretreatment with dexamethasone (DEX), not during or post-DOX treatment. DEX influence appears to be mediated via glucocorticoid receptor and enhances cardiomyocyte-specific gene expression. Therefore, for the gen- eral assessment of cytotoxicity, non-genetically engineered ESC-derived cardiomyocytes are sufficient but for the molecular assessment of DOX-induced toxicity, genetically engineered purified ESC-derived cardiomyocytes are necessary. © 2012 Elsevier Ltd. All rights reserved. 1. Introduction The importance of embryonic stem cells (ESCs) in drug toxicity is well established (Mcneish, 2004; Buesen et al., 2009; Whitlow et al., 2007). ESCs are either used directly for teratology or differentiated to different cell types including cardiomyocytes, hepatocytes, and neurons (Farokhpour et al., 2009; Baharvand et al., 2006; Meamar et al., 2010). ∗ Corresponding author at: Department of Cell and Molecular Biology, Royan Insti- tute for Animal Biotechnology, P.O. Box 815896-8433, Isfahan, Iran. Tel.: +98 311 26129003; fax: +98 311 2605525. ∗∗ Corresponding author at: Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, Iran. Tel.: +98 21 22306485; fax: +98 21 22310406. E-mail addresses: mh nasr@royaninstitute.org (M.H. Nasr-Esfahani), Baharvand@royaninstitute.org (H. Baharvand). Doxorubicin (DOX), an anthracycline antibiotic, is one of the most effective anti-tumor agents used in the treatment of human malignancies. Unfortunately a major side effect of DOX treatment is cardiotoxicity (Chen et al., 2007; Minotti et al., 2004; Bristow et al., 1978), which may be ameliorated by corticosteroid ther- apy (Sun et al., 2008; Farokhpour et al., 2009; Morrissy et al., 2010). This cytoprotective effect is believed to be associated with increased expression of the bcl-xL gene that inhibits caspase activ- ity (Chen et al., 2005). We have previously shown that pretreatment with 10 M dexamethasone (DEX) maintained the beating func- tions of DOX-treated ESCs-derived cardiomyocytes (Farokhpour et al., 2009). However, one of the drawbacks of that study was the presence of a heterogeneous population of cells which impeded the proper assessment of biological characteristics such as dose response curve, the inability to discriminate apoptosis between cardiomyocytes and other cell populations, and the assessment of gene expression directly related to cardiomyocytes. Therefore, the current study was undertaken to evaluate the role of genetically versus non-genetically engineered ESCs in the evaluation of drug 0040-8166/$ – see front matter © 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.tice.2012.09.009