• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Research for the in vitro reconstruction of a hepatic tissue and for the treatment of hepatic failure by hepatic tissue transplantation

Research Project

Project/Area Number 12470243
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General surgery
Research InstitutionSapporo Medical University

Principal Investigator

MOCHIZUKI Yohichi  Sapporo Medical University, Cancer Research Institute, Professor, 附属がん研究所, 教授 (40045381)

Co-Investigator(Kenkyū-buntansha) KOJIMA Takashi  Sapporo Medical University, School of medicine, Assistant Professor, 附属がん研究所, 教授 (30260764)
MITAKA Toshihiro  Sapporo Medical University, Cancer Research Institute, Assistant Professor, 附属がん研究所, 助教授 (50231618)
HIRATA Koichi  Sapporo Medical University, School of medicine, Professor, 医学部, 教授 (50136959)
TAKEDA Hiroshi  Sapporo Medical University, Cancer Research Institute, Instructor, 附属がん研究所, 助手 (00333310)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 2001: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2000: ¥9,100,000 (Direct Cost: ¥9,100,000)
KeywordsSmall hepatocyte / Progenitor cell / Bone marrow interstitial cells / Maturation / Extracellular matrix / Scaffold / Human hepatocyte / Cryopreservation / 増殖 / 分化 / 移植 / 類肝組織 / ミニ肝臓
Research Abstract

Small hepatocytes (SHs), which are known to be hepatic progenitor cells, were isolated from an adult rat liver. We found methods by which SHs could differentiate into mature hepatocytes and form three-dimensional structures. SHs in a colony sometimes change their shape from small to large and from flat to rising/piled-up. Although this morphological change occurs when hepatic nonparenchymal cells such as stellate cells invade under SH colonies and produce an extracellular matrix, an overlay of Matrigel^<TM> on SHs can induce similar phenomena. The differentiated SHs express hepatocyte nuclear factor (HNF) 4, HNF6, and CCAAT/enhancer binding protein α, which are expressed in mature hepatocytes. In addition, not only albumin and transferrin but also carbamoylphosphate synthetase (CPS) I and glutamine synthetase, which are key enzymes of urea synthesis and ammonia metabolism, are also expressed. Furthermore, isozymes of cytochrome P450 can be induced by specific drugs. On the other hand, SH colonies could be cryopreserved for more than 6 months and the maximum period of the storage was about 90 weeks. About 60% of SH colonies survive and attached on the dishes. The surviving SHs could proliferate and the average area of SH colonies was about 7.5 times larger at day 15 than at day 1. Albumin production increased with time in culture. In addition, the cells produced other serum proteins such as transferrin and fibrinogen, and expressed CPS I and tryptophan 2,3-dioxygenase.
Human hepatocytes were isolated and we found the proliferation of SHs in the culture. In addition, when the cells were cultured in a collagen sponge, the proliferation of biliary epithelium was observed and mature hepatocytes were maintained with three-dimensional structures for a long time.
We found that interstitial cells derived from rat bone marrow could support the maintenance of differentiated functions of primary hepatocytes.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (72 results)

All Other

All Publications (72 results)

  • [Publications] 三高 俊広: "肝臓における幹細胞システムと肝組織構築(総説)"実験医学. 18(4). 431-437 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toru Mizuguchi, et al.: "Paper is compatible bed for rat hepatocytes"Artificial Organs. 24(4). 271-277 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Masato Isobe, et al.: "Role of inducible nitric oxide synthetase on hepatic ischemia and reperfusion injury"Transplantation Proceedings. 32(7). 1650-1652 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tadashi Katsuramaki, et al.: "Changes in hepatic venous oxygen saturation in hepatic warm ischemia/renerfusion injury in pigs"Surgery Today. 30(4). 343-351 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Minoru Nagayama, et al.: "Changes in hepatic microdialysate uric acid levels in porcine liver ischemia/reperfusion"Transplantation Proceedings. 32(7). 2282-2284 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tadashi Katsuramaki, et al.: "Different changes of endothelin-1 after reperfusion in a warm ischemia/reperfusion and transplantation model in pig liver"Transplantation Proceedings. 32(7). 2276-2278 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takashi Kojima, et al.: "Occludin and claudin-1 concerntrate in the midbody of immortalzed mouse hepatocytes during cell division"Journal of Histochemistry and Cytochemistry. 49(3). 333-340 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takashi Kojima, et al.: "Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes"Experimental Cell Research. 263(2). 193-201 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 三高俊広: "肝臓の幹細胞(総説)"Molecular Medicine. 38(1). 8-15 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 三高俊広, 他: "肝臓の再生(総説)"肝胆膵. 42(2). 195-201 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toshihiro Mitaka: "Hepatic Stem Cells : From Bone Marrow to Hepatocytes (Review)"Biochemical Biophysical Research Communication. 281(1). 1-5 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Koichi Hirata, et al.: "Sepsis and cholestasis : basic findings in the sinsoid and bile canaliculus (Review)"Journal of Hepatobiliary Pancreatic Surgery. 8(1). 20-26 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 三高俊広, 他: "肝臓の幹細胞システム(総説)"GI Research. 19(3). 229-234 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nagato Higaki, et al.: "Maitenance of connexin 32 and 26 expression in primary cultured rat hepatocytes treated with 3-acetylpyridine"Journal of Gastroenterology and Hepatology. 16(7). 806-815 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toru Mizuguchi, et al.: "Regulation of c-met expression in rats with acute hepatic failure"Journal of Surgical Research. 99(2). 385-396 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toru Mizuguchi, et al.: "Enhanced proliferation and differentiation of rat hepatocytes cultured with bone marrow stromal cells"Journal of Cellular Physiology. 189(1). 106-119 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toshihiro Mitaka, et al.: "Expression of carbamoylphosphate synthetase I and glutamine synthetase in hepatic organoids reconstructed by rat small hepatocytes and hepatic nonparenchymal cells"Cell and Tissue Research. 306(3). 467-471 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 三高俊広: "動き出した再生医療の臨床〜肝臓〜"分子細胞治療. 1(1). 78-85 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toru Mizuguchi, et al.: "Effects of bone marrow stromal cells on the structural and functional polarity of primary rat hepatocvtes"In Vitro Cellular and Developmental Biology Animal. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nagato Katsura, et al.: "Long-term culture of primary human hepatocytes with preservation of proliferative capacity and differentiated functions"Journal of Surgical Research. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shinichiro Ikeda, et al.: "Proliferation of rat small hepatocytes after long-term cryopreservation"Journal of Hepatology. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toshihiro Mitaka: "Reconstruction of hepatic organoid by hepatic stem cells"Journal of Hepatobiliary Pancreatic Surgery. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toshihiro Mitaka, et al.: "The Hepatocyte Review : Chapter 14"Kluwer Academic Publishers (Dordrecht, The Netherlands). 624 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 池田 慎一郎, 他: "幹細胞・クローン研究プロトコール:第5章 肝前駆細胞からの増殖と分化:小型肝細胞(small hepatocyte)の分離方法"羊土社、東京. 254 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mizuguchi T, et al.: "Paper is compatible bed for rat hepatocytes"Artif. Organs. 24(4). 271-277 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] MitakaT, et al.: "Chapter 14: Small Hepatocytes in Primary Cultures"In: Berry M. and Edward AM, eds. The Hepatocyte Review, Kluwer academic publishers, Dordrecht. 195-208 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Isobe M, et al.: "Role of inducible nitric oxide synthetase oh hepatic ischemia and reperfusion injury"Transplant Proc. 32(7). 1650-1652 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katsuramaki T, et al.: "Changes in hepatic venous oxygen saturation in hepatic warm ischemia/reperfusion injury in pigs."Surg Today. 30 (4). 343-351 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nagayama M, et al.: "Changes in hepatic microdialysate uric accid levels in porcine liver ischemia/reperfusion"Transplant Proc. 32 (7). 2282-2284 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katsuramaki T, et ai.: "Different changes of endothelin- 1 after reperfusion in a warm ischemia/reperfusion and transplantation model in pig liver."Transplant Proc. 32 (7). 2276-2278 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kojima T, et al.: "Occludin and claudin- 1 concentrate in the midbody of immortalized mouse hepatocytes during cell division"J. Histochem. Cytochem.. 49(3). 333-339 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kojima T, et al.: "Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes"Exp. Cell Res.. 263 (2). 193-201 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitaka T.: "Hepatic Stem Cells: From Bone Marrow Cells to Hepatocytes"Biochem. Biophys. Res. Commun. 281(1). 1-5 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hirata K, et al.: "Sepsis and eholestasis: basic findings in the sinusoid and bile canaliculus"J Hepatobiliary Pancreat Surg.. 8(1). 20-26 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Higaki N, et al.: "Maintenance of connexin 32 and 26 expression in primary cultured rat hepatocytes treated with 3- acetylpyridine"J Gastroenterol Hepatol.. 16(7). 806-815 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mizuguchi T, et al.: "Regulation of c-met expression; in rats with acute hepatic failure"J. Surg. Res.. 99(2). 385-396 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mizuguchi T, et al.: "Enhanced proliferation and differentiation of rat hepatocytes cultured with bone marrow stromal cells"J. Cell Physiol.. 189(1). 106-119 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitaka T.: "Expression of carbamoylphosphate synthetase I and glutamine synthetase in hepatic organoids reconstructed by rat small hepatocytes and hepatic nonparehchymal cells"Cell Tissue Res.. 306(3). 467-471 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mizuguchi T, et al.: "Effects of bone marrow stromal cells on tne structural and functional polarity of primary rat heptocytes."In Vitro Cell Dev. Biol.. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Katsura N, et al.: "Long-Term Culture of Primary Human Hepatoqytes vyith Preservation of Proliferatiye Capacity and Differentiated Functions"J Surg Res. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ikeda S., et al.: "Proliferation or rat small hepatocytes alter long-term cryopreservation"J Hepatol.. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitaka T.: "Reconstruction ol hepatic organoia by hepatic stem cells"J Hepatobiliary Pancreat Surg.. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 三高俊広: "肝臓における幹細胞システムと肝組織構築(総説)"実験医学. 18(4). 431-437 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toru Mizuguchi: "Paper is compatible bed for rat hepatocytes"Artificial Organs. 24(4). 271-277 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Masato Isobe, et al.: "Role of inducible nitric oxide synthetase on hepatic ischemia and reperfusion injury"Transplantation Proceedings. 32(7). 1650-1652 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tadashi Katsuramaki, et al.: "Changes in hepatic venous oxygen saturation in hepatic warm ischemia/reperfusion injury in pigs"Surgery Today. 30(4). 343-351 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Minoru Nagayama, et al.: "Changes in hepatic microdialysate uric acid levels in porcine liver ischemia/reperfusion"Transplantation Proceedings. 32(7). 2282-2284 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tadashi Katsuramaki, et al.: "Different changes of endothel in-1 after reperfusion in a warm ischemia/reperfusion and transplantation model in pig liver"Transplantation Proceedings. 32(7). 2276-2278 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] 三高俊広: "肝臓の再生(総説)"肝胆膵. 42(2). 195-201 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takashi Kojima et al.: "Occludin and claudin-1 concerntrate in the midbody of immortalzed mouse hepatocytes during cell division"Journal of Histochemistry and Cytochemistry. 49(3). 333-340 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takashi Kojima. et al.: "Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes"Experimental Cell Research. 263(2). 193-201 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Koichi Hirata, et al.: "Sepsis and cholestasis : basic findings in the sinusoid and bile canaliculus. (Review)"Journal of Hepatobiliary Pancreatic Surgery. 8(1). 20-26 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toshihiro Mitaka, et al.: "Hepatic Stem Cells ; From Bone Marrow to Hepatocytes (Review)"Bicohemical and Biophysical Research Communication. 281(1). 1-5 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 三高 俊広, 他: "肝臓の幹細胞(総説)"Molecular Medicine. 38(1). 8-15 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 三高俊広, 他: "肝臓の幹細胞kステム(総説)"GI Research. 19(3). 229-234 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nagato Higaki et al.: "Maitenance of connexin 32 and 26 expression in primary cultured rat hepatocytes treated with 3-acetylpyridine"Journal of Gastroenterology and Hepatology. 16(7). 806-815 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toru Mizuguchi, et al.: "Regulation of c-met expression in rats with acute hepatic failure"Journal of Surgical Research. 99(2). 385-396 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toru Mizuguchi et al.: "Enhanced proliferation and differentiation of rat hepatocytes cultured with bone marrow stromal cells"Journal of Cellular Physiology. 189(1). 106-109 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toshihiro Mitaka, et al.: "Expression of carbamoylphosphate synthetase 1 and glutamine synthetase in hepatic organoids reconstructed by rat small hepatocytes and hepatic nonparenchymal cells"Cell and Tissue Research. 306(3). 467-471 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toru Mizuguchi et al.: "Effects of bone marrow stromal cells on the structural and functional polarity of primary rat hepatocytes"In Vitro Cellular and Developmental Biology Animal. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 三高俊広: "動き出した再生医療の臨床〜肝臓〜"分子細胞治療. 1(1). 78-85 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nagato Katsura et al.: "Long-term culture of primary human hepatocytes with preservation of proliferative capacity and differentiated functions"Journal of Surgical Research. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shinichiro Ikeda, et al.: "Proliferation of rat small hepatocytes after long-term cryopreservation"Journal of Hepatology. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Toshihiro Mitaka, et al.: "The Hepatocyte Review : Chapter 14"Kluwer Academic Publishers (Dordrecht, The Netherlands). 624 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] 池田慎一郎, 他: "幹細胞・クローン研究プロトコール:第5章 肝前駆細胞からの増殖と分化:小型肝細胞(small hepatocyte)の分離方法"羊土社、東京. 254 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 三高俊広: "肝臓における幹細胞システムと肝組織構築"実験医学. 18(4). 431-437 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mizuguchi T,Mitaka T, et al.: "Paper is compatible bed for rat hepatocytes."Artificial Organs. 24(4). 271-277 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 三高俊広: "肝臓の幹細胞"Molecular Medicine. 38(1). 8-15 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kojima T,Kokai Y,Mochizuki Y, et al.: "Cx32 but not Cx26 is associated with tight junctions in primary cultures of rat hepatocytes"Experimental Cell Research. 263(2). 193-201 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mitaka T,: "Hepatic Stem Cells : From Bone Marrow Cells to Hepatocytes."Biochemical Biophysical Research Communication. 281(1). 1-5 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 三高俊広,池田慎一郎,杉本真一: "肝臓の再生."肝胆膵. 42(2). 195-201 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mitaka T and Sato F.: "The Hepatocyte Review, Chapter 14."Kluwer Academic Publishers (Dordrecht, The Netherlands). 624 (2000)

    • Related Report
      2000 Annual Research Report

URL: 

Published: 2000-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi