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Department of Radiology, The Third Affiliated Hospital of Sun Yat‐sen University, Guangzhou, China Hepatocytes from human bone marrow‐derived mesenchymal stem cells (hBM‐MSCs) are expected to be a useful source for cell transplantation. However, relatively low efficiency of hepatic differentiation of hBM‐MSCs remains unsolvable in clinical application. Hepatocyte nuclear factor 4alpha (HNF4α), a critical transcription factor, is essential for the entire process of liver development. The purpose of this study was to construct MSCs with over expression HNF4α and investigate their hepatic differentiation and therapeutic potential for treating Fulminant hepatic failure (FHF) rats, and track their survival and biodistribution after transplantation by bioluminescence imaging (BLI). HNF4α gene was transduced into hBM‐MSCs by lentiviral vector (pLV/Final‐puro‐hHNF4α‐hrGFP). HNF4α‐trans‐duced MSCs (E7‐hHNF4α) and GFP‐transduced MSCs cells (E7‐hrGFP ) were labeled with pLENTi‐CMV‐luc2‐mKate2 and analyzed for their hepatic functions such as measurements of albumin, urea, glucose, cytochrome P450 activity,Indocy‐anine green (ICG) uptake and release, and drug metaboliza‐tion in vitro.

FHF modals of Sprague‐Dawley (SD) rats were established and divided into three groups: PBS, E7‐hrGFP and E7‐hHNF4α cells' transplantation. After 2.0×10 6 cells transplantation, BLI was used to dynamically track the survival and biodistribution of implanted E7‐hrGFP cells and E7‐hHNF4α cells. The restoration of biological functions of the livers receiving transplantation was assessed via a variety of approaches such as mortality rate determination, serum biochemical analysis, and histological, immunohistochemical, and genetic analysis. In vitro, E7‐hHNF4α cells showed mature hepatic functions including albumin secretion, urea production, glycogen storage, cytochrome P450 activity, ICG uptake and release and drug metabolization. They improved liver functions in vivo after transplantation into the D‐galactosamine‐injured rats' liver as evidenced by the fact that AST, ALT, TBIL returned to normal levels in recipient FHF rats.