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Objective Drug-induced hepatotoxicity is often caused by cytochrome P450 (CYP)-dependent metabolism of drugs into reactive metabolites.Assessment of hepatotoxicity induced by bioactive compounds is hampered by low CYP expression within in vitro cell lines.To overcome this limitation,piggyBac transposition and monoclonal expansion were used to generate a HepG2 cell line with stable and homogenously high expression of CYP3A4,a prominent CYP isoform.Our studies demonstrate the generated lines constant CYP3A4 expression and activity for over 40 cell passages;to date,it has been in subculture for more than a year without addition of Puromycin.