Toxicity of ecstasy (MDMA) towards embryonic stem cell-derived cardiac and neural cells Rokhsareh Meamar a,b , Fereshteh Karamali a , Hamid Mirmohamad Sadeghi b , Mahmood Etebari b , Mohammad Hossein Nasr-Esfahani a,** , Hossein Baharvand c,d, * a Department of Cell and Molecular Biology, Royan Institute for Animal Biotechnology, P.O. Box 815896-8433, Esfahan, Iran b Department of Pharmacology, School of Pharmacy and Pharmaceutical Sciences, University of Isfahan, Esfahan, Iran c Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, Iran d Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran article info Article history: Received 30 August 2009 Accepted 10 March 2010 Available online 15 March 2010 Keywords: Embryonic stem cells Cardiomyocyte differentiation Neuronal differentiation Ecstasy MDMA abstract ‘‘Ecstasy” or methylenedioxymethamphetamine (MDMA) is primarily a recreational drug commonly used during the child bearing period, thus, there is a major concern regarding the embryonic and fetal toxicity of this drug. Here, we report the cardio- and neuro-toxic effects of MDMA on beating embryoid bodies (EBs) and neural cell-containing EBs derived from mouse embryonic stem cell (ESCs). Based on our linear discriminate function, MDMA is considered to be a moderate or weak teratogen. Moreover, the genera- tion of EBs with neural cell morphology and the expression of MAP2, a mature neuron marker, decrease more when MDMA is administered during the EB formation stage rather than post-plated EBs. In addi- tion, the ID50 (inhibition of differentiation) of EBs with neural cell morphology is less than cardiac cells. In conclusion, MDMA causes a marked reduction in beating cardiomyocytes and neurons in ESC cultures, and this drug has a more potent toxicity on neural rather than cardiac cell differentiation. Ó 2010 Elsevier Ltd. All rights reserved. 1. Introduction The recreational drug 3,4-methylenedioxymetamphetamine (MDMA; ecstasy) is popular among young adults, including child- bearing women (Ho et al., 2004). MDMA has been shown to pro- duce both cardiotoxicity and neurotoxicity in humans and animals. Acute administration of MDMA increases heart rate, blood pressure and myocardial oxygen consumption in humans (Lester et al., 2000; O’Cain et al., 2000) and animals (Bexis and Docherty, 2006) which may ultimately result in hypertension, arrhythmias, cardiac ischemia and heart failure. Even a single administration of MDMA can produce significant cardiovascular toxicity in vivo (Cerretani et al., 2008). On the other hand, it is suggested that ingestion of MDMA by a pregnant female would result in abnormal neural and cardiovascular development in the fetus (Colado et al., 1997), since this drug with its low molecular weight and high lipo- solubility readily crosses the placenta (Meyer et al., 2004). More- over, MDMA-induced damage to neurons, which lasts for months in rats and years in primates, has been demonstrated (Bowyer et al., 2003; Green et al., 2003; Hatzidimitriou et al., 1999). Similar studies indicate that MDMA-induced neurodegeneration may also occur in human users abusers (McCann et al., 1998). These studies primarily suggest that in vivo neurotoxicity of MDMA is mainly in- duced by the release of dopamine and serotonin which cause an excitotoxic response that includes glutamate release and increased intracellular calcium, resulting in apoptosis and neural death. A recent study by Kindlundh-Hogberg et al., has also reported the neurotoxicity of MDMA to neural precursor cells in vitro (Kindlundh-Hogberg et al., 2009). MDMA is primarily a recreational drug commonly used during the child bearing period, thus, there is a major concern regarding embryos exposed to this drug during the embryonic period from conception to 12 weeks or the fetal period from 12 weeks until birth. Although the overall toxicity of MDMA is well accepted, however, the consequences of MDMA abuse at the DNA level (mutagenesis and/or DNA integrity), cellular proliferation (cyto- toxicity), cellular determination, specification and differentiation (cell lineage specification), the integration of the latter two phe- nomena (embryo toxicity) as well as post differentiation or post organogenesis (fetal toxicity) and their eventual birth defects (teratology) have not been fully characterized. Therefore, there is an increasing need to integrate embryotoxicity tests to evaluate the effects of novel chemical and pharmaceutical agents on the 0887-2333/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.tiv.2010.03.005 * Corresponding author at: Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, Iran. Tel.: +98 21 22306485; fax: +98 21 22310406. ** Corresponding author. Tel.: +98 311 26129003; fax: +98 311 2602555. E-mail addresses: mh.nasr-esfahani@RoyanInstitute.org (M.H. Nasr-Esfahani), Baharvand50@yahoo.com, Baharvand@RoyanInstitute.org (H. Baharvand). Toxicology in Vitro 24 (2010) 1133–1138 Contents lists available at ScienceDirect Toxicology in Vitro journal homepage: www.elsevier.com/locate/toxinvit