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. 2018 Apr 5;9:296. doi: 10.3389/fphar.2018.00296

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Copyright © 2018 Bencsik, Kupai, Görbe, Kenyeres, Varga, Pálóczi, Gáspár, Kovács, Weber, Takács, Hajdú, Fabó, Cseh, Barna, Csont, Csonka, Dormán and Ferdinandy.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Experimental protocol for cell culture studies and for the ex vivo rat heart model of AMI. (A) Isolated neonatal rat cardiac myocytes were subjected to 4 h of simulated ischemia followed by 2 h of simulated reperfusion. At the end of the reperfusion, cell viability was determined by using calcein flurescence. (B) Isolated adult rat hearts were perfused according to Langendorff and a 30-min global, no-flow ischemia was applied after a 20 min equilibration period. Subsequently, 2 h reperfusion was applied and then infarct size was determined. The hearts were perfused with Krebs-Henseleit solution containing lead candidates or vehicle from 20 min prior to the global ischemia until the 60th min of reperfusion.