Functional repolarization of reconstructed hepatic organoid : Pumping function of bile canaliculi
スポンサーリンク
概要
- 論文の詳細を見る
The morphogenesis and movement of bile canaliculi (BC) are not well understood. Once hepatocytes are isolated from the liver, they rapidly lose their polarized function, and BC disappear. We found that small hepatocytes (SHs) could proliferate and form a large colony. Some colonies changed their shapes from flat to piled-up with time in culture. The piled-up cells show the morphology of mature hepatocytes (MHs) and BC-like structures can be observed between the piled-up cells. We investigated whether piled-up cells had membrane polarity, how BC-like structures contracted, and whether these structures were functionally active as BC. Hepatic cells including SHs, were isolated from the adult rat liver and cultured. Immunocytochemistry was carried out for BC proteins such as ectoATPase, 5'-nucleotidase (5'NT), and multidrug-resistance associated protein 2 (MRP2), and for gap junctional protein, connexin 32 (Cx32). Furthermore, phase-contrast microscopy with time-lapse equipment was used for the observation of BC contractions and the transfer of microinjected dye. Secretion of fluorescein into the reconstructed BC was also observed. Immunocytochemistry for BC proteins revealed that these proteins localized in intercellular space, coinciding with BC-like structures formed between piled-up cells. Cx32-immunoreactive dots were observed in the basolateral membranes of piled-up cells. Fluorescein diacetate was added to the medium and metabolized fluorescein was secreted into BC and accumulated without leakage. Time-lapse images showed that continuous contractions of reconstructed BC occurred in a coordinated manner and a unidirectional flow was observed. In conclusion, we showed that SHs could differentiate into highly polarized hepatocytes and reconstruct a hepatic organoid, and that the polarized cells could acquire the ability to transport bile by pumping.
- 一般社団法人日本機械学会の論文
著者
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Ikeda Mariko
School Of Fundamental Sci. And Technol. Keio Univ.
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Ikeda Mariko
Department Of Chemistry And Life Science Nihon University
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Kobayashi Hirosuke
School Of Allied Health Sciences Kitasato Univ.
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Ikeda Mariko
Department Of System Design Engineering Keio University
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Kohara Hiroshi
Department Of Pathophysiology Cancer Research Institute Sapporo Medical University School Of Medicin
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Ikeda Mariko
Department Of Applied Chemistry. Faculty Of Engineering Toyama University
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Tanishita Kazuo
Department of System Design Engineering Keio University
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MITAKA Toshihiro
Department of Pathology, Cancer Research Institute, Sapporo Medical University School of Medicine
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Sudo Ryo
Department of System Design Engineering, Keio University
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Sudo Ryo
Massachusetts Institute of Technology, Biological Engineering Division
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Sudo Ryo
Biological Engineering Division M.i.t.
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Kohara Hiroshi
Department Of Anesthesiology Sapporo Medical University School Of Medicine
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Tanishita Kazuo
Department Of System Design Engineering Keio Univ.
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Mitaka Toshihiro
Department Of Molecular Pathophysiology Cancer Research Institute Sapporo Medical University School
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Tanishita Kazuo
Department Of System Engineering
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Mitaka Toshihiro
Department Of Pathophysiology Cancer Reserch Institute Sapporo Medical University School Of Medicine
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Tanishita Kazuo
Department Of System Design Engineering
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