Hepatocyte nuclear factor Foxa-2 effectively differentiates mesenchymal stem cells to functional hepatocytes

Abstract

Overexpression of hepatocyte nuclear factor (Foxa-2), a master regulator significantly enhances the hepatic differentiation by triggering targeted liver-specific gene expression for liver development. We investigated the function of key regulators that facilitate hepatic differentiation. The functional hepatocyte was determined by observing morphological changes, expression of liver specific markers through immunocytochemical analysis, qRT-PCR, Western Blot and specific liver function assays. Our study confirmed that induction of functional hepatocyte shows typical epithelial morphology, express hepatic genes and acquire hepatocyte functions. Notably, these cells expressed the markers of mature hepatocytes, including albumin, tyrosine aminotransferase, cytokeratin, Cyp7A1 and hepatic transcription factors. Furthermore, these cells exhibited hepatic functions in vitro, including glycogen storage, albumin production, urea secretion and functional assessment of hepatocytes. We have developed a robust and efficient method to differentiate mesenchymal stem cells into induced functional hepatocytes, which exhibits characteristics of mouse hepatocytes. Apparently, induction of functional hepatocytes will pave a better cellular therapy or effective cell source within a short timeline to facilitate the development of hepatocytes for future clinical applications of regenerative medicine.