A novel 3D liver organoid system for elucidation of hepatic glucose metabolism

Authors

  • Yanhua Lu,

    1. Department of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, Zhejiang 310027, China; telephone: 86-571-87953193; fax: 86-571-87951227
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  • Guoliang Zhang,

    1. Institute of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, Zhejiang 310012, P.R. China
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  • Chong Shen,

    1. Department of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, Zhejiang 310027, China; telephone: 86-571-87953193; fax: 86-571-87951227
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  • Korkut Uygun,

    1. Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, Boston, Massachusetts
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  • Martin L. Yarmush,

    1. Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, Boston, Massachusetts
    2. Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey
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  • Qin Meng

    Corresponding author
    1. Department of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, Zhejiang 310027, China; telephone: 86-571-87953193; fax: 86-571-87951227
    • Department of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, Zhejiang 310027, China; telephone: 86-571-87953193; fax: 86-571-87951227.
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Abstract

Hepatic glucose metabolism is a key player in diseases such as obesity and diabetes as well as in antihyperglycemic drugs screening. Hepatocytes culture in two-dimensional configurations is limited in vitro model for hepatocytes to function properly, while truly practical platforms to perform three-dimensional (3D) culture are unavailable. In this work, we present a practical organoid culture method of hepatocytes for elucidation of glucose metabolism under nominal and stress conditions. Employing this new method of culturing cells within a hollow fiber reactor, hepatocytes were observed to self-assemble into 3D spherical organoids with preservation of tight junctions and display increased liver-specific functions. Compared to both monolayer culture and sandwich culture, the hepatocyte organoids displayed higher intracellular glycogen content, glucose consumption, and gluconeogenesis and approached the in vivo values, as also confirmed by gene expression of key enzymes. Moreover, hepatocyte organoids demonstrated more realistic sensitivity to hormonal challenges with insulin, glucagon, and dexamethasone. Finally, the exposure to high glucose demonstrated toxicities including alteration of mitochondrial membrane potential, lipid accumulation, and reactive oxygen species formation, similar to the in vivo responses, which was not captured by monolayer cultures. Collectively, hepatocyte organoids mimicked the in vivo functions better than hepatocyte monolayer and sandwich cultures, suggesting suitability for applications such as antihyperglycemic drugs screening. Biotechnol. Bioeng. 2012; 109:595–604. © 2011 Wiley Periodicals, Inc.

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