2. Academic Validation
  3. Generation of canine gallbladder cholangiocyte organoids from healthy and gallbladder mucocele patient dogs for functional analysis of anion channels

Generation of canine gallbladder cholangiocyte organoids from healthy and gallbladder mucocele patient dogs for functional analysis of anion channels

  • Vet J. 2025 Oct:313:106417. doi: 10.1016/j.tvjl.2025.106417.
Itsuma Nagao 1,  Hirotaka Tomiyasu 2,  Yuko Goto-Koshino 1,  Kota Nakase 1,  Takumi Tsuruta 1,  Yuto Namiki 1,  Ryuichiro Sato 3,  Yu Takahashi 4,  Yuko Hashimoto 5,  Yosuke Takahashi 5,  Naohiro Takahashi 6,  James K Chambers 6,  Kazuyuki Uchida 6,  Kenji Baba 7,  Yoko M Ambrosini 8,  Masaru Okuda 1
Affiliations

Affiliations

  • 1 Laboratory of Veterinary Internal Medicine, Graduate School of Agricultural and Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
  • 2 Laboratory of Veterinary Internal Medicine, Graduate School of Agricultural and Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: atomi@g.ecc.u-tokyo.ac.jp.
  • 3 Nutri-Life Science Laboratory, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
  • 4 Food Biochemistry Laboratory, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
  • 5 Veterinary Medical Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
  • 6 Laboratory of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
  • 7 Laboratory of Veterinary Internal Medicine, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
  • 8 Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, 255 E Main St, Pullman, WA 99163, United States.
PMID: 40835125 DOI: 10.1016/j.tvjl.2025.106417
Abstract

Gallbladder mucocele (GBM) is one of the most common gallbladder disorders in dogs. Recent studies have suggested a potential association between GBM development and reduced expression of anion channel-related molecules in gallbladder epithelial cells. However, further investigation has been limited due to the lack of an effective in vitro culture system to validate the functions of the molecules. This study aimed to establish gallbladder cholangiocyte organoids (GCOs) from canine gallbladders and evaluate anion channel functions. GCOs were generated from gallbladders of three clinically healthy dogs and three GBM-affected dogs, followed by morphological evaluation, comparisons of anion channel-related gene expressions, and functional assessment of anion channels, specifically cystic fibrosis transmembrane conductance regulator (CFTR) and calcium-activated chloride channels (CaCCs), using Organoid swelling assays. Most GCOs derived from normal gallbladder (N-GCOs) exhibited a round, cystic morphology, whereas GCOs from GBM-affected gallbladder (GBM-GCOs) predominantly displayed a budding morphology with narrowed lumen. Although no significant differences in anion channel-related gene expression were observed between the two groups, the functions of both CFTR and CaCCs were significantly impaired in GBM-GCOs. The morphological differences between the groups might be attributed to the reduced ion secretions and subsequent shortages of water transport into the Organoid lumen in GBM-GCOs, leading to insufficient lumen expansion. These findings highlighted a potential link between anion channel dysfunction and GBM pathogenesis. Further studies utilizing canine GCOs are warranted to deepen our understanding of the role of anion channel dysfunction in GBM development.

Keywords

Anion channel; CFTR; Calcium-activated chloride channel; Dog; Gallbladder cholangiocyte organoid; Gallbladder mucocele.

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