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Establishment of Biliary Epithelial Cell Lines from the Hamster

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Hepatobiliary and Pancreatic Carcinogenesis in the Hamster

We describe a method for the simultaneous culturing of biliary epithelial cells (BECs) from the gallbladder (GB), extrahepatic bile duct (EBD), and intrahepatic bile duct (IBD) of hamsters. The GB, EBD and IBD were excised from the biliary tree after collagenase perfusion of the liver. These biliary segments were minced into fragments, which were embedded in collagen gel and cultured in Dulbeccos Modified Eagle Medium/HamF12 Medium containing 10% fetal bovine serum. The various cells subsequently spread from the fragments and formed cellular sheets. After the fragments and flattened cells were removed under phase-contrast microscopy, the sheets remaining were found to be composed of cuboidal cells. These cuboidal cells expressed gamma glutamyl transpeptidase and cytokeratin 7, which are known to be specific markers of BECs. Ultrastructurally, there were many microvilli on the luminal surface and junctional complex and interdigitation was identifiable on the lateral surfaces. BEC cultures were subcultured by digestion with collagenase and dispase and then dissociated by subsequent digestion in trypsin and ethylenediaminetetraacetic acid. They were maintained in collagen gel for up to 8 weeks. After several passages, the BECs in the culture eventually grew and showed vacuoles in the cytoplasm. They demonstrated irreversible growth arrest at 9 weeks. The BECs tended to form cystic structures when they were transplanted with collagen gel into the interscapular fat pads of the syngeneic hamsters.

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Authors and Affiliations

  1. Department of Surgery, National Hospital Organization Saga National Hospitl, 1-20-1 Hinode, Saga, 849-0923, Japan

    Takayuki Asakawa

  2. Department of Surgery, National Hospital Organization Nagasaki Medical Center, 2-1001-1 Kubara, Omura, Nagasaki, 856-8562, Japan

    Amane Kitasato

  3. Department of Surgery, Nagasaki Yurino Hospital, 1155-2 Motomura, Togitsu, Nishisonogi, Nagasaki, 851-2103, Japan

    Tsutomu Tomioka

  4. Department of Surgery, Nagasaki Prefectural Shimabara Hospital, 7895 Shimokawajiri, Shimabara, Nagasaki, 855-0861, Japan

    Ryuji Tsutsumi

Authors
  1. Takayuki Asakawa
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  2. Amane Kitasato
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  3. Tsutomu Tomioka
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  4. Tamotsu Kuroki
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  5. Ryuji Tsutsumi
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  6. Yoshitsugu Tajima
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  7. Takashi Kanematsu
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Editors and Affiliations

  1. Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki, 852-8501, Japan

    Yoshitsugu Tajima M.D., Ph.D. , Tamotsu Kuroki M.D., Ph.D.  & Takashi Kanematsu M.D., Ph.D. ,  & 

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Asakawa, T. et al. (2009). Establishment of Biliary Epithelial Cell Lines from the Hamster. In: Tajima, Y., Kuroki, T., Kanematsu, T. (eds) Hepatobiliary and Pancreatic Carcinogenesis in the Hamster. Springer, Tokyo. https://doi.org/10.1007/978-4-431-87773-8_14

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