Role of Nrf2 Signaling in Development of Hepatocyte-Like Cells

Abstract

Generation of hepatocytes from human adipose-derived mesenchymal stem cells (hADSCs) could be a promising alternative source of human hepatocytes. However, mechanisms to differentiate hepatocytes from hADSCs are not fully elucidated. In this study, we investigated the role of nuclear factor erythroid-2 related factor 2 (Nrf2) in differentiation of hepatocyte-like cells (HLCs). We used our established three-step differentiation protocol to develop HLCs from hADSCs. Significant nuclear translocation of Nrf2 occurred from day 11 (Step 2) until the end of HLC differentiation. There were no significant differences in Nrf2 translocation rates among the four experimental groups (activin-A, GSK3 inhibitor, Nrf2 siRNA, and control) at day 6 (end of Step 1). Nuclear translocation of Nrf2 in the GSK3 inhibitor-treated group was obviously higher than the other groups at day 11 (Step 2). Moreover, nuclear translocation of Nrf2 in the GSK3 inhibitor-treated group was notably higher than the other groups during Step 3. CYP3A4 activity (Luciferin-IPA assay) of the GSK3 inhibitor-treated group was significantly higher than the other three groups. Nrf2 was activated during differentiation of HLCs, and inhibition of Nrf2 delayed maturation and impaired the function of HLCs. Thus, Nrf2 might be a notable target for developing highly functional human HLCs.