Bile Canalicular Formation in Hepatic Organoid Reconstructed by Rat Small Hepatocytes and Nonparenchymal Cells

Ryo Sudo, Shinichiro Ikeda, Shinichi Sugimoto, Keisuke Harada, Koichi Hirata, Kazuo Tanishita, Yohichi Mochizuki, Toshihiro Mitaka

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The morphogenesis and movement of bile canaliculi (BC) are not well understood. This is because culture of hepatocytes that maintain polarity of cell membranes and possess highly differentiated functions has never been successful. We found that small hepatocytes (SHs), which are known to be hepatic progenitor cells, could proliferate and differentiate into mature hepatocytes and that BC-like structures developed between rising/piled-up cells. We investigated how BC-like structures developed with maturation of SHs and whether the structures were functionally active as BC. Hepatic cells, including SHs, were isolated from an adult rat liver and cultured. Immunocytochemistry and immunoblotting for BC proteins, such as ectoATPase, 5′-nucleotidase, dipeptidylpeptidase IV, and multidrug-resistance associated protein 2, were examined and time-lapse microscopy was used for the observation of BC contractions. Secretion of bilirubin into the reconstructed BC was also observed. The results of immunocytochemistry, immunoblots, and immunoelectron micrographs revealed that BC proteins were localized in the intercellular space that coincided with BC-like structures reconstructed between rising/piled-up cells. Tight junction-associated protein ZO-1 was also expressed along the BC-like structures. Bilirubin added to the medium were secreted into BC-like structure and accumulated without leakage. Time-lapse microscopy showed continuous contractions of reconstructed BC. In conclusion, BC-like structures reconstructed by SHs may be functional with membrane polarity, secretory ability, and motility. These results show that this culture system may suitable for investigating the mechanism of the formation of BC and their functions.

Original languageEnglish
Pages (from-to)252-261
Number of pages10
JournalJournal of Cellular Physiology
Volume199
Issue number2
DOIs
Publication statusPublished - 2004 May
Externally publishedYes

Fingerprint

Bile Canaliculi
Organoids
Bile
Rats
Hepatocytes
Liver
Bilirubin
Microscopic examination
5'-Nucleotidase
Proteins
Cell membranes
Cell culture
Microscopy
Immunohistochemistry
Zonula Occludens-1 Protein
Membranes
Extracellular Space

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Sudo, R., Ikeda, S., Sugimoto, S., Harada, K., Hirata, K., Tanishita, K., ... Mitaka, T. (2004). Bile Canalicular Formation in Hepatic Organoid Reconstructed by Rat Small Hepatocytes and Nonparenchymal Cells. Journal of Cellular Physiology, 199(2), 252-261. https://doi.org/10.1002/jcp.10407

Bile Canalicular Formation in Hepatic Organoid Reconstructed by Rat Small Hepatocytes and Nonparenchymal Cells. / Sudo, Ryo; Ikeda, Shinichiro; Sugimoto, Shinichi; Harada, Keisuke; Hirata, Koichi; Tanishita, Kazuo; Mochizuki, Yohichi; Mitaka, Toshihiro.

In: Journal of Cellular Physiology, Vol. 199, No. 2, 05.2004, p. 252-261.

Research output: Contribution to journalArticle

Sudo, R, Ikeda, S, Sugimoto, S, Harada, K, Hirata, K, Tanishita, K, Mochizuki, Y & Mitaka, T 2004, 'Bile Canalicular Formation in Hepatic Organoid Reconstructed by Rat Small Hepatocytes and Nonparenchymal Cells', Journal of Cellular Physiology, vol. 199, no. 2, pp. 252-261. https://doi.org/10.1002/jcp.10407
Sudo, Ryo ; Ikeda, Shinichiro ; Sugimoto, Shinichi ; Harada, Keisuke ; Hirata, Koichi ; Tanishita, Kazuo ; Mochizuki, Yohichi ; Mitaka, Toshihiro. / Bile Canalicular Formation in Hepatic Organoid Reconstructed by Rat Small Hepatocytes and Nonparenchymal Cells. In: Journal of Cellular Physiology. 2004 ; Vol. 199, No. 2. pp. 252-261.
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