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Treatment of liver failure by transplantation of hepatocytes derived from mouse and monkey ES cells and human umbilical cord blood

Research Project

Project/Area Number 16390357
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General surgery
Research InstitutionTokyo medical and Dental University

Principal Investigator

TERAMOTO Kenichi  Tokyo medical and Dental University, surgery, Assistant professor, 大学院・医歯学総合研究科, 助教授 (80197813)

Co-Investigator(Kenkyū-buntansha) ARII Shigeki  Tokyo medical and Dental University, surgery, professor, 大学院・医歯学総合研究科, 教授 (50151171)
TANAKA Yujiro  Tokyo medical and Dental University, professor, 医学部付属病院, 教授 (70236644)
SAITO Keiko  Tokyo medical and Dental University, instructor, 難治疾患研究所, 教務職員 (50178969)
ASAHINA Kinji  Tokyo medical and Dental University, Assistant, 難治疾患研究所, 助手 (40345294)
TERAOKA Hirobumi  Tokyo medical and Dental University, M.R.I, professor, 難治疾患研究所, 教授 (30019137)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥13,900,000 (Direct Cost: ¥13,900,000)
Fiscal Year 2005: ¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 2004: ¥7,300,000 (Direct Cost: ¥7,300,000)
KeywordsCynomolgus / Embryonic stem cells / Hepatocyte differentiation / Transgenic mouse / umbilical cord blood / liver failure / hepatocyte transplantation / teratoma / マウスES細胞 / 胚様体 / 肝細胞分化 / 細胞移植 / 肝細胞
Research Abstract

1) Monkey embryonic stem (ES) cells have characteristics that are similar to human ES cells, and might be useful as a substitute model for preclinical research. When embryoid bodies (EBs) formed from monkey ES cells were cultured, expression of many hepatocyte-related genes including cytochrome P450 (Cyp) 3a and Cyp7a1 was observed. Hepatocytes were immunocytochemically observed using antibodies against albumin (ALB), cytokeratin-8/18, and a1-antitrypsin in the developing EBs. The in vitro differentiation potential of monkey ES cells into the hepatic lineage prompted us to examine the transplantability of monkey EB cells. As an initial approach to assess the repopulation potential, we transplanted EB cells into immunodeficient urokinasetype plasminogen activator transgenic mice that undergo liver failure. After transplantation, the hepatocyte colonies expressing monkey ALB were observed in the mouse liver. Fluorescence in-situ hybridization revealed that the repopulating hepatocytes ar … More ise from cell fusion between transplanted monkey EB cells and recipient mouse hepatocytes. In contrast, neither cell fusion nor repopulation of hepatocytes was observed in the recipient liver after undifferentiated ES cell transplantation. These results indicate that the differentiated cells in developing monkey EBs, but not contaminating ES cells, generate functional hepatocytes by cell fusion with recipient mouse hepatocytes, and repopulate injured mouse liver.
2) We previously reported that hepatocytes can be differentiated from embryonic stem (ES) cells by way of embryoid body (EB) formation and are transplantable into the mouse liver. However, the transplantation of EB-derived cells frequently resulted in teratoma formation in the recipient liver. In the present study, we eliminated the tumorigenic cells from EB outgrowths and examined the effects of enriched ES-cell-derived hepatocyte transplantation into an injured liver. On day 15 in culture, the EBs were partially disaggregated and subcultured. Hepatocytes in the subcultured cells were examined by the expression of hepatocyte markers. Undifferentiated cells contaminating in the EB-derived cells were eliminated by Percoll discontinuous gradient centrifugation. Furthermore, undifferentiated cells, endothelial cells, and macrophages were eliminated by magnetic cell sorting using platelet/endothelial cell adhesion molecule (PECAM)-1 and Mac-1 antibodies. These enriched ES-cell-derived hepatocytes were then transplanted into the injured mouse liver. Percoll centrifugation and PECAM-1 antibodies eliminated the undifferentiated cells expressing Oct-3/4 from the EB-derived cells. ES-cell-derived hepatocytes showed expression of liver-related genes, synthesis of urea and glycogen, and structural characteristics during subculture. A transplantation study showed that the enriched ES-cell-derived hepatocytes integrated into the injured mouse liver and produced no teratomas. When the ES-cell-derived hepatocytes were transplanted into a CCl4-injured liver, the liver function was subsequently improved. Functional hepatocytes can be differentiated from mouse ES cells by way of EB formation. The elimination of undifferentiated cells from the EBs provides transplantable cells for liver failure without tumorigenicity. Less

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report

Research Products

(19 results)

All 2006 2005 2004

All Journal Article (19 results)

  • [Journal Article] -生体防御機構の解明と最先端医療への道-5ES細胞からの肝細胞分化誘導2006

    • Author(s)
      寺本研一, 神代祐至, 入江工, 有井滋樹, 朝比奈欣時治, 斉藤佳子, 寺岡弘文
    • Journal Title

      別冊サイトプロダクション 別冊

      Pages: 31-35

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Hepatocyte differentiation from ES cells.2006

    • Author(s)
      K Teramoto, Y Kumashiro, T Irie, S Arii, K Asahina, K Shimizu-Saito, H Teraoka
    • Journal Title

      Cytoprotection (Separate volume)

      Pages: 31-35

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Generation of hybrid hepatocytes by cell fusion from monkey embryoid body cells in the injured mouse liver2006

    • Author(s)
      Kentaro Okamura, Kinji Asahina, Hiroaki Fujimori, Rie OZeki, Keiko Shimizu-Saito, Yujiro Tanaka, Kenichi Teramoto, Shigeki Arii, Kozo Takase, Miho Kataoka, Yoshinori Soeno, Chise Tateno, Katsutoshi Yoshizato, Hirobumi Teraoka
    • Journal Title

      Histochemistry and Cell Biology 125(3)

      Pages: 245-257

    • NAID

      120000876692

    • Related Report
      2005 Annual Research Report
  • [Journal Article] -生体防御機構の解明と最先端医療への道-ES細胞からの肝細胞分化誘導2006

    • Author(s)
      寺本研一, 神代祐至, 入江工, 有井滋樹, 朝比奈欣時治, 斉藤佳子, 寺岡弘文
    • Journal Title

      別冊サイトプロダクション (別冊)

      Pages: 31-35

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Teratoma formation and hepatocyte differentiation in mouse liver transplanted with mouse embryonic stem cell-derived embryoid bodies.2005

    • Author(s)
      K.Teramoto, Y.Hara, Y.Kumashiro, R.Chinzei, Y.Tanaka, K.Shimizu-Saito, K.Asahina, H.Teraoka, S.Arii
    • Journal Title

      Transplantation Proceedings 37(1)

      Pages: 285-286

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Isolation of Hepatocyte-like Cells From Mouse Embryoid Body Cells.2005

    • Author(s)
      Y Kumashiro, K Teramoto, Shimizu-Saito, K Asahina, H Teraoka, S Arii
    • Journal Title

      Transplantation Proceedings 37(1)

      Pages: 299-300

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Enrichment of Hepatocytes Differentiated from Mouse Embryonic Stem Cells as a Transplantable Source.2005

    • Author(s)
      Yuji Kumashiro, Kimji Asahina, Rie Ozeki, Keiko Shimizu-Saito, Yujiro Tanaka, Yujiro Kida, Kouji Inoue, Michinari Kaneko, Tetsuji Sato, Kenichi Teramoto, Shigeki Arii, Hirobumi Teraoka
    • Journal Title

      Transplantation 79(5)

      Pages: 550-557

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Hepatocyte differentiation from embryonic stem cells and umbilical cord blood cells.2005

    • Author(s)
      K.Teramoto, K Asahina, Y Kumashiro, S Kakinuma, R.Chinzei, KShimizu-Saito, Y Tanaka, H Teraoka, S Arii
    • Journal Title

      Joulnal of Hepatobiliary Pancreat Suregery 12

      Pages: 196-202

    • NAID

      10016432279

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Generation of hybrid hepatocytes by cell fusion from monkey embryoid body cells in the injured mouse liver.2005

    • Author(s)
      Kentaro Okamura, Kinji Asahina, Hiroaki Fujimori, Rie Ozeki, Keiko Shimizu-Saito, Yujiro Tanaka, Kenichi Teramoto, Shigeki Arii, Kozo Takase, Miho Kataoka, Yoshinori Soeno, Chise Tateno, Katsutoshi Yoshizato, Hirobumi Teraoka
    • Journal Title

      Histochemistry and Cell Biology 125(3)

      Pages: 245-257

    • NAID

      120000876692

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Teratoma formation and hepatocyte differentiation in mouse liver transplanted with mouse embryonic stem cell-derived embryoid bodies.2005

    • Author(s)
      K.Teramoto, Y.Hara, Y.Kumashiro, R.Chinzei, Y.Tanaka, K.Shimizu -Saito, K.Asahina, H.Teraoka, S.Arii
    • Journal Title

      Transplantation Proceedings 37(1)

      Pages: 285-286

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Hepatocyte differentiation from embryonic stem cells and umbilical cord blood cells.2005

    • Author(s)
      K.Teramoto, K Asahina, Y Kumashiro, S Kakinuma, R.Chinzei, K Shimizu-Saito, Y Tanaka, H Teraoka, S Arii
    • Journal Title

      Journal of Hepatobiliary Pancreat Surgery 12

      Pages: 196-202

    • NAID

      10016432279

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Teratoma formation and hepatocyte differentiation in mouse liver transplanted with mouse embryonic stem cell-derived embryoid bodies2005

    • Author(s)
      K.Teramoto, Y.Hara, Y.Kumashiro, R.Chinzei, Y.Tanaka, K.Shimizu-Saito, K.Asahina, H.Teraoka, S.Arii
    • Journal Title

      Transplantation Proceedings 37(1)

      Pages: 285-286

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Enrichment of Hepatocytes Defferentiated from Mouse Embryonic Stem Cells as a Transplantable Source2005

    • Author(s)
      Yuji Kusashiro, Kimji Asahina, Rie Ozeki, Keiko Shimizu-Saito, Yujiro Tanaka, Yujiro Kida, Kouji Inoue, Michinari Kaneko, Tetsuji Sato, Kenichi Teramoto, Shigeki Arii, Hirobumi Teraoka
    • Journal Title

      Transplantation 79(5)

      Pages: 550-557

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Hepatocyte differentiation from embryonic stem cells and umbilical cord blood cells2005

    • Author(s)
      KENICHI TERAMOTO, KINJI ASAHINA, YUJI KUMASHIRO, SEI KAKINUMA, RYOKO CHINZEI, KEIKO SHIMIZU-SAITO, YUJIRO TANAKA, HIROBUMI TERAOKA, SHIGEKI ARII
    • Journal Title

      Joulnal of Hepatobiliary Pancreat Suregery 12

      Pages: 196-202

    • NAID

      10016432279

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Teratoma Formation and Hepatocyte Differentiation in Mouse Liver Transplanted with Mouse Embryonic Stem Cell-Derived Embryoid Bodies2005

    • Author(s)
      K.Teramoto, Y.Hara, Y.Kumashiro, R.Chinzei, Y.Tanaka, K.Shimizu-Saito, K.Asahina, H.Teraoka
    • Journal Title

      Transplantation Proceedings (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Expression of the liver-specific gene Cyp7a1 reveals hepatic differentiation in embryoid bodies derived from mouse embryonic stem cells.2004

    • Author(s)
      Kinji Asahina, Hiroaki Fujimori, Keiko Shimizu-saito, Yuji Kumashiro, Kentaro Okamura, Yujiro Tanaka, Kenichi Teramoto, Shigeki Arii, Hirobumi Teraoka
    • Journal Title

      Genes to Cells 9

      Pages: 1297-1308

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 臍帯血有核細胞による肝臓再生医療の可能性2004

    • Author(s)
      朝比奈 欣治, 齋藤 佳子, 寺岡 弘文, 柿沼 晴, 寺本 研一
    • Journal Title

      細胞 36(2)

      Pages: 16-19

    • Related Report
      2004 Annual Research Report
  • [Journal Article] ES細胞からの肝細胞分化の試み2004

    • Author(s)
      寺本研一, 神代祐至, 入江工, 有井滋樹
    • Journal Title

      低温医学 30(2)

      Pages: 32-36

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Expression of the liver-specific gene Cyp7al reveals hepatic differentiation in embryoid bodies derived from mouse embryonic stem cells2004

    • Author(s)
      Kinji Asahina, Hiroaki Fujimori, Keiko Shimizu-saito, Yuji Kumashiro, Kentaro Okamura, Yujiro Tanaka, Kenichi Teramoto, Shigeki Arii, Hir
    • Journal Title

      Genes to Cells 9

      Pages: 1297-1308

    • Related Report
      2004 Annual Research Report

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Published: 2004-03-31   Modified: 2016-04-21  

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