Wei-Lun Tsai1,2, Po-Hung Yeh3,4, Chia-Yun Tsai5,6, Chin-Tsung Ting7,8, Yen-Hui Chiu9, Mi-Hua Tao10, Wan-Chun Li5,6,9, Shih-Chieh Hung3,4,10,11,12,13. 1. Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan. 2. Medical School, National Yang-Ming University, Taipei, Taiwan. 3. Stem Cell Laboratory, Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan. 4. Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan. 5. Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan. 6. Department of Dentistry, School of Dentistry, National Yang-Ming University, Taipei, Taiwan. 7. Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan. 8. Division of Gastrointestinal Surgery, Department of Surgery, Ren-Ai Branch, Taipei City Hospital, Taipei, Taiwan. 9. Department of Education and Research, Taipei City Hospital, Taipei, Taiwan. 10. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. 11. Institute of Clinical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan. 12. Integrative Stem Cell Center, Department of Orthopedics, China Medical University Hospital, Taichung, Taiwan. 13. Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan.
Abstract
BACKGROUND AND AIM: In view of its unique properties of detoxification and involvement of metabolic and biochemical functions, in vitro hepatocyte culture serves as a valuable material for drug screening and mechanistic analysis for pathology of liver diseases. The restriction of rapid de-differentiation and inaccessibility of human hepatocytes from routine clinical procedure, however, limits its use. METHODS: To address this issue, the effort to direct human mesenchymal stem cells (hMSCs) into hepatocytes using a modified protocol was proposed. With the additional treatment of histone deacetylase inhibitor (HDACi) and DNA methyltransferase inhibitor (DNMTi), in vitro hMSC-derived hepatocytes were cultivated and their hepatic characteristics were examined. RESULTS: By using a modified protocol, it was shown that Trichostatin A and 5-aza-2-deoxycitidine protected differentiating cells from death and could sufficiently trigger a wide range of liver-specific markers as well as liver functions including albumin production, glycogen storage, and urea cycle in hMSC-derived hepatocytes. The increased mRNA expression for hepatitis C virus (HCV) entry including CD81, Occludin, LDL receptor, and scavenger receptor class B type I in hMSC-derived hepatocytes was also detected, implying its potential to be utilized as an in vitro model to analyze dynamic HCV infection. CONCLUSIONS: The present study successfully established a protocol to direct hMSCs into hepatocyte-like cells suggesting the beneficial impact to apply HDACi and DNMTi as potent modulators for hMSCs to liver differentiation.
BACKGROUND AND AIM: In view of its unique properties of detoxification and involvement of metabolic and biochemical functions, in vitro hepatocyte culture serves as a valuable material for drug screening and mechanistic analysis for pathology of liver diseases. The restriction of rapid de-differentiation and inaccessibility of human hepatocytes from routine clinical procedure, however, limits its use. METHODS: To address this issue, the effort to direct human mesenchymal stem cells (hMSCs) into hepatocytes using a modified protocol was proposed. With the additional treatment of histone deacetylase inhibitor (HDACi) and DNA methyltransferase inhibitor (DNMTi), in vitro hMSC-derived hepatocytes were cultivated and their hepatic characteristics were examined. RESULTS: By using a modified protocol, it was shown that Trichostatin A and 5-aza-2-deoxycitidine protected differentiating cells from death and could sufficiently trigger a wide range of liver-specific markers as well as liver functions including albumin production, glycogen storage, and urea cycle in hMSC-derived hepatocytes. The increased mRNA expression for hepatitis C virus (HCV) entry including CD81, Occludin, LDL receptor, and scavenger receptor class B type I in hMSC-derived hepatocytes was also detected, implying its potential to be utilized as an in vitro model to analyze dynamic HCV infection. CONCLUSIONS: The present study successfully established a protocol to direct hMSCs into hepatocyte-like cells suggesting the beneficial impact to apply HDACi and DNMTi as potent modulators for hMSCs to liver differentiation.
Authors: Paula Mueller; Markus Wolfien; Katharina Ekat; Cajetan Immanuel Lang; Dirk Koczan; Olaf Wolkenhauer; Olga Hahn; Kirsten Peters; Hermann Lang; Robert David; Heiko Lemcke Journal: Cells Date: 2020-02-22 Impact factor: 6.600