Our previous work revealed that a polyethyleneimine (PEI)-based gene delivery causes robust and sustained expression of exogenous genes in human adipose-derived stem cells (hADSCs). Here we use this method to test whether a single introduction of cDNAs for the three cardiomyogenic reprogramming genes (GATA4, MEF2C, and TBX5) might be sufficient to induce transdifferentiation of hADSCs towards the cardiomyogenic lineage. A single transfection results in sustained expression of the introduced genes for more than two weeks. hADSCs exhibit undetectable or very low levels of mRNAs for endogenous GATA4, MEF2C and TBX5. However, mRNAs for these endogenous factors become apparent at similar to 2 weeks after transfection and keep increasing until the end of experimental period at the fifth week. Concordant with these cardiomyogenic genes, Nkx2.5 mRNA becomes significant at similar to 2 weeks and gradually increases until the end of experimental period. Several other cardiomyogenic mRNAs were also significant at 5 weeks. Thus, a single transfection of cDNAs for the cardiomyogenic reprogramming genes using a PEI based method induces transdifferentiation of ADSCs. (C) 2016 Elsevier Inc. All rights reserved.