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In liver, drug metabolism related CYP enzymes release depend on sex hormone, but most studies on new drug development for treatment of liver failure did not considered this phenomenon. In recent era of stem cells, there has been a growing interest in generation of stem cell based hepatocyte like cells for treatment of liver diseases or development of new drug. In this study we analyzed, the effect of gender in induced Hepatocyte like cells (miHep) using male or female mouse embryonic fibroblast (MEF). For generation of miHep, male and female MEF were co-transfected with 3 lentiviral vectors (MOI=1) inserted each liver transcription factors (hHnf1a, hHnf4a, hFox) for 2 days and cultured in hepatocyte induction medium, continuously. After one month, clusters of hepatocyte like cells with polygonal cytoplasm and bipolar nucleic acid were clearly observed and continuously increased the number of hepatocyte like cells clusters as time goes on, regardless of their gender. After 3 months, expression of exo- and endogenic transcription factors identified by real time PCR and confirmed expression of not only 3 endogenic- but also 3 exogenic transcription factors. For transcriptome profiling, generated miHep along with positive and negative control groups, healthy liver and MEF of male and female mice from the same strain, were subjected to RNA analysis. Transcriptomes of generated miHeps were similar to transcriptomes of liver tissue, especially drug metabolism, glycogen metabolism, epithelial-mesenchymal transition (EMT), cell adhesion molecules (CAM) and hepatocyte genes. Overall, transcriptomes of all CYP enzymes were normalized with female, and revealed that female liver showed 2-fold up change of CYP genes than male; however, these results were not found in miHeps. Based on above results, we concluded that there is no gender effect on miHeps generation and on in vitro culturing without sex hormone. These results can be utilized in future study of cell therapy and cell line generation in drug development for liver failure.This work was conducted with the support of “Cooperative Research Program for Agriculture Science &Technology Development” carried out with the support of Program (no. PJ01100203).