title: Enhanced hepatogenic transdifferentiation of human adipose tissue mesenchymal stem cells by gene engineering with Oct4 and Sox2.
abstract: Adipose tissue mesenchymal stem cells (ATMSCs) represent an attractive tool for the establishment of a successful stem cell-based therapy in the field of liver regeneration medicine. ATMSCs overexpressing Oct4 and Sox2 (Oct4/Sox2-ATMSCs) showed enhanced proliferation and multipotency. Hence, we hypothesized that Oct4 and Sox2 can increase "transdifferentiation" of ATMSCs into cells of the hepatic lineage. In this study, we generated Oct4- and Sox2-overexpressing human ATMSCs by liposomal transfection. We confirmed the expression of mesenchymal stem cell surface markers without morphological alterations in both red-fluorescent protein (RFP) (control)- and Oct4/Sox2-ATMSCs by flow cytometry. After induction of differentiation into hepatocyte-like cells, the morphology of ATMSCs changed and they began to appear as round or polygonal epithelioid cells. Hepatic markers were evaluated by reverse transcription-polymerase chain reaction and confirmed by immunofluorescence. The results showed that albumin was strongly expressed in hepatogenic differentiated Oct4/Sox2-ATMSCs, whereas the expression level of α-fetoprotein was lower than that of RFP-ATMSCs. The functionality of hepatocytes was evaluated by periodic acid-Schiff (PAS) staining and urea assays. The number of PAS-positive cells was significantly higher and urea production was significantly higher in Oct4/Sox2-ATMSCs compared to that in RFP-ATMSCs. Taken together, the hepatocyte-like cells derived from Oct4/Sox2-ATMSCs were mature hepatocytes, possibly functional hepatocytes with enhanced capacity to store glycogen and produce urea. In this study, we demonstrated the enhanced transdifferentiation of Oct4- and Sox2-overexpressing ATMSCs into hepatocyte-like cells that have enhanced hepatocyte-specific functions. Therefore, we expect that Oct4/Sox2-ATMSCs may become a very useful source for hepatocyte regeneration or liver cell transplantation.
authors: Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Sei-Myoung
authors: SM
authors: Han
authors: Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Ye-Rin
authors: YR
authors: Coh
authors: Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Jin-Ok
authors: JO
authors: Ahn
authors: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Goo
authors: G
authors: Jang
authors: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Soo Young
authors: SY
authors: Yum
authors: Stem Cell Research Center, K-STEMCELL Co. Ltd., Seoul, 153-768, Republic of Korea.
authors: Sung-Keun
authors: SK
authors: Kang
authors: Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Hee-Woo
authors: HW
authors: Lee
authors: Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea; Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Republic of Korea.
authors: Hwa-Young
authors: HY
authors: Youn
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D060846:Cell Engineering; D054338:Cell Transdifferentiation / Q000235:genetics*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D008164:Luminescent Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D050814:Octamer Transcription Factor-3 / Q000235:genetics*; D055748:SOXB1 Transcription Factors / Q000235:genetics*; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D000097573:Red Fluorescent Protein