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Development of the packed-bed type bioartificial liver

Research Project

Project/Area Number 12470251
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Digestive surgery
Research InstitutionUniversity of Tsukuba

Principal Investigator

YANAGI Kennichi  Institute of Basic Medical Sciences, Lecturer, 基礎医学系, 講師 (70239797)

Co-Investigator(Kenkyū-buntansha) TAKADA Yasutsugu  University of Kyoto, School of Medicine, Associate professor, 医学部, 助教授 (10272197)
MIYOSHI Hirotoshi  Institute of Basic Medical Sciences, Assistant professor, 基礎医学系, 講師 (70292547)
OHSHIMA Norio  Institute of Basic Medical Sciences, Professor, 基礎医学系, 教授 (50015971)
KANAMORI Toshiyuki  National Institute of Advanced Industrial Science and Technology, Institute for Materials and Chemical Process, Chief Researcher, 物質プロセス研究部門, 主任研究官
TANIGUCHI Hideki  Yokohama City University, School of Medicine, Professor, 医学部, 教授 (70292555)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥13,900,000 (Direct Cost: ¥13,900,000)
Fiscal Year 2002: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2001: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2000: ¥5,700,000 (Direct Cost: ¥5,700,000)
KeywordsBioartificial liver / Porcine / Fulminant hepatic failure / Packed-bed type reactor / α-amanitine / lipopolisaccharide / High-density culture of hepatocytes / Ammonium / 生存時間 / 動物モデル / 充填層型リアクタ / ポリビニルホルコール樹脂多孔質体 / 肝細胞
Research Abstract

The authors have developed a packed-bed type bioreactor using porous polyvinyl formal resin as cell supporting material tor use as a bioartificial liver. Large scale and high density culture of hepatocytes was achieved using the reactor. In the present study, the reactor volume was scaled up from a 10 cm^3 of the prototype to 1,000 cm^3, and the metabolic performance of the reactor was evaluated in in vitro culture experiments and ex vivo perfusion experiments using a fulminant hepatic failure model of a porcine.
[Methods] A PVF resin with a mean pore size of 250 μm cut into cubes (2 mm or 3 mm) was used as a cell supporting material. Packed-bed reactors with the culture volume 1,000 cm^3 were used. Hepatocytes obtained from pigs were inoculated onto the PVF resin. Culture medium (Williams' E medium supplemented with 10% fetal bovine serum, hormones and antibiotics) was perfused from the reservoir into the reactor through an oxygenator. Oxygen concentration and pH were measured at the o … More utlet of the reactor, and were maintained at 40% and 7.4, respectively. In in vitro culture experiments performed for a week, metabolic activities of the cultured hepatocytes were evaluated in terms of a few liver-specific functions, i.e., ammonium metabolism, urea synthesis, lidocaine metabolism and albumin secretion. The immobilized cell density was measured by the DNA quantity in the PVF resin cubes. Morphology of the immobilized hepatocytes was observed. In ex vivo perfusion experiments, acute hepatic failure was induced by injecting alpha-amanitine (0.15 mg/kg) and lipopolysaccharide (1 μg/kg) in the portal vein of pigs (8 - 12 kg). Twenty-two hours after injection of the drugs, porcine blood was dialyzed against the perfusate of the packed-bed type reactor using a hollow-fiber type dialyzer. Blood samples were drawn periodically for later analysis. Extracorporeal perfusions using the module without hepatocytes were performed as a control.
[Results and Conclusion] Hepatocytes were well immobilized in the PVF. The maximum cell count in the reactor reached a value of 1.2 x 10^<10> cells (120 g) on Day 2. The cell density gradually decreased with culture time. Hepatocytes immobilized in the packed-bed reactor showed satisfactory and stable metabolic activities comparable to those in vivo liver. Morphological observation revealed that the immobilized cells showed well-differentiated morphologies similar to those observed in vivo. In ex vivo perfusion experiments, following the administration of α-amanitine and LPS, pigs showed a diseased state of fulminant hepatic failure, and died at an average of 50 hours after injection of the drugs. In the pigs that received hepatic support from the bioartificial liver, the rise in the blood ammonium concentration was suppressed, and there was a tendency for an extension in survival times. In conclusion, the packed-bed type bioreactor is a promising module of importance in developing a bioartificial liver. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] 柳 健一他: "充填層型バイオ人工肝臓の開発-培養ブタ肝細胞の機能と形態の評価(学会抄録)"医用電子と生体工学. 39. 675 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Yanagi et al.: "Development of a packed-bed type bioartificial liver -Metabolic performance of cultured porcine hepatocytes utilizing a scaled-up bioreactor"Tissue Engineering for Therapeutic Use 6 eds ; Ikada Y, Umakoshi Y and Hotta T. 67-73 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Yanagi, N.Ohshima: "Improvement of metabolic performance of cultured hepatocytes by high oxygen tension in the atmosphere"Artificial Organs. 25(1). 1-6 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳 健一他: "バイオ人工肝臓への応用を目的とする充填層型リアクタの開発-リアクタの性能改善とスケールアップ-"肝臓. 41(3). 76-77 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳 健一他: "充填層型バイオ人工肝臓の開発◇リアクタのスケールアップと性能評価◇(学会抄録)"人工臓器. 29(3). s-142 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 大島宣雄他: "再生医工学の手法によるバイオ人工肝臓の開発(学会抄録)"医用電子と生体工学. 38(特別号). 5 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] N.Ohshima et al.: "Tissue-engineered bioartificial organs that utilize 3-dimensional cell culture scaffold of a highly porous polymer material (meeting abstract)"医用電子と生体工学. 38(特別号). 26 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳健一他: "ブタの薬剤性劇症肝不全モデルの作成(学会抄録)"生体医工学. 40・特別号. 104 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳健一他: "充填層型バイオ人工肝臓の開発 in vitroおよびex vivoの性能評価(学会抄録)"人工臓器. 31・2. s-10 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi, K., et al.: "Extracorporeal liver assist for pigs of drug-induced acute hepatic failure using a packed-bed type bioartificial liver (meeting abstract)"Tissue Engineering. 8(6). 1221 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳健一他: "充填層型バイオ人工肝臓によるブタ薬剤性劇症肝不全の治療効果(学会抄録)"再生医療-日本再生医療学会雑誌. 2・増刊号. 90 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳健一他: "充填層型バイオ人工肝臓によるブタ薬剤性劇症肝炎の治療効果(学会抄録)"日本外科学会雑誌. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi, K., et al.: "Extracorporeal liver assist for pigs with drug induced fulminant hepatic failure using a packed-bed type bioartificial liver (meeting abstract)"ASAIO J. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi, K., et al.: "Performance of a scaled-up packed-bed reactor for use as a bioartificial liver (meeting abstract)"ASAIO J. 46(2). 226 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Improvement of metabolic performance of cultured hepatocytes by high oxygen tension in the atmosphere"Artificial Organs. 25(1). 1-6 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Performance of a scaled-up packed-bed reactor for use as a bioartificial liver (meeting abstract)"ASAIOJ. 46(2). 226 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Extracorporeal liver assist for pigs of drug-induced acute hepatic failure using a packed-bed type bioartificial liver (meeting abstract)"Tissue Engineering. 8(6). 1221 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Extracorporeal liver assist for pigs with drug induced fulminant hepatic failure using a packed-bed type bioartificial liver (meeting abstract)"ASAIOJ. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Development of a packed-bed reactor for use as a bioartificial liver (in Japanese)"Kanzo (Acta Hepatologica Japonica). 41(3). 76-77 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Development of a packed-bed type bioartificial liver : Performance of the scaled-up reactor (meeting abstract in Japanese)"Jinko-zoki (The Japanese Journal of Artificial Organs). 29(3). s142 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ohshima,N., et al.: "Tissue-engineered bioartificial organs that utilize 3-dimensional cell culture scaffold of a highly porous polymer material (meeting abstract in Japanese)"Iyodennshi-to-seitaikogaku (Japanese journal of medical electronics and biological engineering). 38. s26 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Development of the packed-bed type bioartificial liver ―Metabolic performance and morphological examination of cultured porcine hepatocytes (meeting abstract in Japanese)"Iyodennshi-to-seitaikogaku (Japanese journal of medical electronics and biological engineering). 39. s675 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al.: "Development of a drug induced fulminant hepatic failure model of a pig (meeting abstract in Japanese)"Iyodennshi-to-seitaikogaku (Japanese journal of medical electronics and biological engineering). 40. s104 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yanagi,K., et al. , eds. by Ikada,Y., Umakoshi,Y. and Hotta,T.: "Development of a packed-bed type bioartificial liver ―Metabolic performance of cultured porcine hepatocytes utilizing a scaled-up bioreactor"Tissue Engineering for Therapeutic Use 6. 67-73 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柳健一: "ブタの薬剤性劇症肝不全モデルの作成(学会抄録)"生体医工学. 40・特別号. 104 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 柳健一: "充填層型バイオ人工肝臓の開発ムin Vitroおよびex vivoの性能評価(学会抄録)"人工臓器. 31・2. s-10 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yanagi, K., et al.: "Extracorporeal liver assist for pigs of drug-induced acute hepatic failure using a packed-bed type bioartificial liver (meeting abstract)"Tissue Engineering. 8・6. 1221 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 柳健一: "充填層型バイオ人工肝臓によるブタ薬剤性劇症肝不全の治療効果(学会抄録)"再生医療-日本再生医療学会雑誌. 2・増刊号. 90 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 柳 健一, 他: "充填層型バイオ人工肝臓の開発-培養ブタ肝細胞の機能と形態の評価"医用電子と生体工学. 39. 675 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Yanagi, et al.: "Development of a packed-bed type bioartificial liver -Metabolic performance of cultured porcine hepatocytes utilizing a scaled-up bioreactor"Tissue Engineering for Therapeutic Use 6. 67-73 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 柳健一 他: "バイオ人工肝臓への応用を目的とする充填層型リアクタの開発-リアクタの性能改善とスケールアップ-"肝臓. 41(3). 76-77 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Yanagi and N.Ohshima: "Improvement of metabolic performance of cultured hepatocytes by high oxygen tension in the atmosphere"Artificial Organs. 25(1). 1-6 (2001)

    • Related Report
      2000 Annual Research Report

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

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