Establishment and application of human embryonic stem cell-derived cardiomyocytes for cardiotoxicity assessment

ZHANG Zhen-zhen; WANG San-long; ZHAO Ya; TANG Wen; ZHANG Ying-li; MA Yue; WANG Ju-feng

Abstract：Objective:To establish human embryonic stem cell-derived cardiomyocytes (hESC-CM) for cardiotoxicity assessment.Methods:Four cardiotoxic drugs were selected as references.They were doxorubicin (anthracycline anticancer drug,0.2,1,5,10 and 20 μmol· L-1),imatinib (tyrosine kinase inhibitor,0.2,1,5,10 and 20 μmol·L-1),isoproterenol (β receptor agonist 0.001,0.01,0.1,1 and 10 μmol·L-1),and verapamil (calcium channel blocker,0.1,0.3,1,3 and 10 μmol· L-1).hESC-CMs were incubated with these drugs.The real-time cell analysis system and high content cell imaging technology were used to monitor the changes in cardiomyocytes' beat rate,amplitude,beat rate regularity index and mitochondrial membrane potential.Potential cardiac toxic of the testing compound A was evaluated too.Results:Beat rate,amplitude,and beat rate regularity,mitochondrial membrane potential of hESC-CMs were significantly decreased after treated with doxorubicin.When hESC-CMs were incubated with 20 μmol·L-1 imatinib,their beat rate,amplitude,beat rate regularity index,and mitochondrial membrane potential were significant inhibited.Isoproterenol caused amplitude fall and beat rate rise,respectively.As increasing incubation time,the effects of isoproterenol were decreased.Treated with verapamil,beat amplitude and beat rate decreased rapidly,the indexes also resumed as time going.There were no any significant changes in beat rate regularity and mitochondrial membrane potential after treatment with isoproterenol and verapamil.When treated with the testing compound A and tofacitinib,beat amplitude,beat rate,rate regularity of hESC-CMs were decreased in a dose-and time-dependent manner.Compound A did not affect mitochondrial membrane potential.Conclusion:The establishment of a feasible cardiotoxicity evaluation model based on hESCCMs in vitro,combined with real-time cell analysis and high content cell imaging technology can be rapid,sensitive,accurate evaluation of drug cardiotoxicity.