Three-Dimensional High-Density Culture of HepG2 Cells in a 5-ml Radial-Flow Bioreactor for Construction of Artificial Liver(Biochemical Engineering)

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概要

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• A three-dimensional high-density cell culture is essential for the construction of an artificial tissue. Many researchers have reported that three-dimensional cell culture enhances cell function. The use of a radial-flow bioreactor (RFB) has enabled the cultivation of cells at high density for constructing a three-dimensional tissue. In this study, we have developed a novel, small RFB, which has a bed volume of 5 ml and is equipped with a porous support as an immobilized scaffold; its performance was tested using the hepatoblastoma cell line, HepG2. Among the other supports tested here, hydroxyl apatite was selected from the viewpoint of its ability to support good cell growth at high density with uniform distribution in a bioreactor. The HepG2 cells grew well in the scaffold under a sufficient supply of nutrients by radial flow and were used to construct a three-dimensional tissue in the scaffold. The concentration of the cells cultivated in this 5-ml RFB reached 10^8 cells/ml and the glucose consumption rate was almost similar to that obtained when using a 30-ml RFB, which has already been reported previously. This high glucose consumption continued over 7 d after the growth phase. Furthermore, albumin production was maintained in the stable phase. Gene expression profiles of cells obtained from long-term cultures in the 5-ml RFB were analyzed. It was found that the expressions of genes encoding the cell cycle-related proteins, cyclins, and cell cycle division 2 (cdc2) were suppressed in the stable phase. In addition, the number of cells incorporating 5'-bromo-2'-deoxyuridine (BrdU) in the stable phase markedly decreased compared with that in the growth phase. These results indicated that the majority of cells in the stable phase remain in the G0/G1 phase. Furthermore, this implies that the three-dimensional tissue constructed in the 5-ml RFB showed the high function similar to a normal liver in the human body. Therefore, the 5-ml RFB was considered as a useful tool and a substitute method for animal experiments.

• 社団法人日本生物工学会の論文
• 2005-03-25

著者

• Honda Hiroyuki

  Department of Biotechnology, School of Engineering, Nagoya University

• OZAWA Shogo

  Project Team for Pharmacogenetics, National Institute of Health Sciences

• Kawamoto Manabu

  Institute Of Rheumatology Tokyo Women's Medical University

• Ozawa Shogo

  Project Team For Pharmacogenetics National Institute Of Health Sciences

• Sawada Junichi

  Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences (nihs)

• OHNO Yasuo

  Division of Pharmacology, National Institute of Health Science

• Kitakaze Masafumi

  Nagoya University Graduate School Of Medicine

• Tomoike Hitonobu

  国立循環器病センター 疫学部

• Tomoike Hitonobu

  国立循環器病センター研究所 疫学部

• Ohe Yuichiro

  国立医薬品食品衛生研究所

• Ohe Yuichiro

  Thoracic Oncology Division National Cancer Center Hospital National Cancer Center

• Ohe Yuichiro

  Department Of Medical Oncology National Cancer Center Hospital

• Ohe Yuichiro

  Department Of Internal Medicine National Cancer Center Hospital

• Ohno Yasuo

  Division Of Pharmacology National Institute Of Health Sciences

• OZAWA Shogo

  Division of Pharmacology, National Institute of Health Sciences

• Manabe Kasumi

  鳥取大学 医学系研究科再生医療学

• Ozawa S

  Project Team For Pharmacogenetics National Institute Of Health Sciences

• Matsuura Kaoru

  国立循環器病センター

• Kobayasi T

  School Of Bioscience And Biotechnology Chubu University

• Sawada Jun-ichi

  Division Of Functional Biochemistry And Genomics National Institute Of Health Sciences

• Tomoike H

  Division Of Preventive Cardiology National Cardiovascular Center

• Ozawa Shogo

  Division Of Pharmacology National Institute Of Health Sciences

• Awata Masaki

  関西ろうさい病院 内科･循環器科

• Ishikawa Yoichi

  エイブル

• Hongo Tomokatsu

  ABLE Corporation

• Kajikawa Mariko

  ABLE Corporation

• Ishida Seiichi

  Division of Pharmacology, National Institute of Health Sciences

• Ishikawa Yoichi

  ABLE Corporation

• UMEZAWA AKIRA

  ABLE Corporation

• KOBAYASHI TAKESHI

  College of Bioscience and Biotechnology, Chubu University

• Kitakaze Masafumi

  Department Of Cardiovascular Medicine National Cardiovascular Center

• Minatoya Kenji

  Department Of Cardiovascular Surgery National Cardiovascular Center

• Ohno Y

  Division Of Pharmacology National Institute Of Health Sciences

• Ohno Y

  Natl. Inst. Health Sci.

• Ohno Yasuo

  Div. Of Pharmacology Biological Safty Research Center National Institute Of Hygienic Sciences

• Ohe Yuichiro

  Division Of Internal Medicine And Thoracic Oncology National Cancer Center Hospital

• Honda Hiroyuki

  Department Of Bioengineering Tokyo Institute Of Technology

• Honda Hiroyuki

  Department Of Biotechnology School Of Engineering Nagoya University

• Ozawa Shogo

  National Institute Of Health Sciences

• Honda Hiroyuki

  Department Of Cardiovascular Research R.i.e.m. Nagoya University

• Ishida Seiichi

  Division Of Pharmacology National Institute Of Health Sciences

• Ishida Seiichi

  National Institute Of Health Sciences

• Ozawa Shogo

  Project Team For Pharmacogenetics National Inst. Of Health Sciences

• Tomoike Hitonobu

  Cardiology And Emergency Medicine National Cardiovascular Center

• Ishikawa Yoichi

  Able Co. Ltd.

• Sawada J

  Mie Univ. School Of Medicine Mie Jpn

• Okada Shuichi

  Hepatobiliary And Pancreatic Oncology Division National Cancer Center Hospital

• Kobayashi Takeshi

  Dep. Of Biological Chemistry Coll. Of Bioscience And Biotechnology Chubu Univ.

• Omori Yoshihito

  Division Of Pharmacology National Institute Of Hygienic Sciences

• Omori Yoshihito

  Division Of Pharmacology Biological Safety Research Center National Institute Of Hygienic Sciences:d

• Ohno Yasuo

  National Inst. Of Health Sci. Tokyo Jpn

• Ohno Yasuo

  Division Of Internal Medicine And Thoracic Oncology National Cancer Center Hospital

• Ohkawa Shinichi

  Division Of Hepatobiliary And Pancreatic Oncology Kanagawa Cancer Center Hospital

• Okamoto Yuko

  Department Of Internal Medicine National Cancer Center Hospital

• Ohe Y

  National Cancer Center Hospital Tokyo

• Kitakaze Masafumi

  First Department Of Medicine And Department Of Pathophysiology Osaka University School Of Medicine

• Kadoma Masaaki

  First Department Of Medicine And Department Of Pathophysiology Osaka University School Of Medicine

• OHKAWA Shinichi

  Kanagawa Cancer Center Hospital

• Honda Hiroyuki

  Department Of Anesthesia Niigata City General Hospital

• Ozawa Shogo

  Department Of Pharmacodynamics And Molecular Genetics School Of Pharmacy Iwate Medical University

• Kobayashi Takeshi

  College Of Bioscience And Biotechnology Chubu University

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