Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems

The pancreatic duct plays a key role in collecting pancreatic juice, which is rich in digestive enzymes. The fluid flows unidirectionally into the duodenum, where it mixes with partially digested food to further facilitate digestion. In this study, we report the generation of pancreatic ductal organ...

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Main Authors: Kosuke Sakaguchi, Chiemi Kimura-Nakajima, Akihiko Inaba, Yoshiko Hatano, Hanako Ogawa, Yuichi Koshiishi, Keisuke Tanaka, Tatsuya Kometani, Makoto Ohmoto, Koji Sato, Hiroo Imai, Ken Iwatsuki
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Cell and Developmental Biology
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Online Access:https://www.frontiersin.org/articles/10.3389/fcell.2025.1593226/full
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author Kosuke Sakaguchi
Chiemi Kimura-Nakajima
Akihiko Inaba
Yoshiko Hatano
Hanako Ogawa
Yuichi Koshiishi
Keisuke Tanaka
Tatsuya Kometani
Makoto Ohmoto
Koji Sato
Hiroo Imai
Ken Iwatsuki
author_facet Kosuke Sakaguchi
Chiemi Kimura-Nakajima
Akihiko Inaba
Yoshiko Hatano
Hanako Ogawa
Yuichi Koshiishi
Keisuke Tanaka
Tatsuya Kometani
Makoto Ohmoto
Koji Sato
Hiroo Imai
Ken Iwatsuki
author_sort Kosuke Sakaguchi
collection DOAJ
description The pancreatic duct plays a key role in collecting pancreatic juice, which is rich in digestive enzymes. The fluid flows unidirectionally into the duodenum, where it mixes with partially digested food to further facilitate digestion. In this study, we report the generation of pancreatic ductal organoids from non-human primates for the first time, aimed at investigating the role of tuft cells that reside in the pancreatic duct since no studies have addressed the role of tuft cells in the pancreas. The organoids were maintained in a medium supplemented with Wnt3a, Noggin, R-spondin, and other factors that support pancreatic duct proliferation. These pancreatic organoids expressed the stem cell marker LGR5 mRNA and the ductal marker protein CK19, although tuft cell markers were not detectable at this stage. Upon stimulation with IL-4/13, tuft cell differentiation was confirmed by immunohistochemistry and transcriptomic analysis. We observed induction of DCLK1, as well as taste signaling molecules such as TRPM5 and PLCβ2, which are markers of type II taste cells. Additionally, upregulation of LYZ and DEFB1 mRNA indicated the expression of antimicrobial peptide markers, alongside molecules associated with inflammation. Furthermore, the differentiated organoids specifically responded to a bitter compound, suggesting that pancreatic tuft cells may play a role in detecting potentially harmful chemicals. Finally, immunohistochemical analysis identified tuft cells in the non-human primate pancreas, supporting their involvement in sensing harmful compounds and regulating protective responses within the pancreas.
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spelling doaj-art-5feb6c09fddc46388ef8a5298bbb47992025-08-20T03:52:14ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-05-011310.3389/fcell.2025.15932261593226Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systemsKosuke Sakaguchi0Chiemi Kimura-Nakajima1Akihiko Inaba2Yoshiko Hatano3Hanako Ogawa4Yuichi Koshiishi5Keisuke Tanaka6Tatsuya Kometani7Makoto Ohmoto8Koji Sato9Hiroo Imai10Ken Iwatsuki11Faculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanGraduate School of Science, Osaka University, Suita, Ōsaka, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanFaculty of Health and Welfare, Takasaki University of Health and Welfare, Takasaki, Gunma, JapanDepartment of Applied Biological Chemistry, The University of Tokyo, Tokyo, JapanCenter for the Evolutionary Origins of Human Behavior, Kyoto University, Kyoto, JapanFaculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, JapanThe pancreatic duct plays a key role in collecting pancreatic juice, which is rich in digestive enzymes. The fluid flows unidirectionally into the duodenum, where it mixes with partially digested food to further facilitate digestion. In this study, we report the generation of pancreatic ductal organoids from non-human primates for the first time, aimed at investigating the role of tuft cells that reside in the pancreatic duct since no studies have addressed the role of tuft cells in the pancreas. The organoids were maintained in a medium supplemented with Wnt3a, Noggin, R-spondin, and other factors that support pancreatic duct proliferation. These pancreatic organoids expressed the stem cell marker LGR5 mRNA and the ductal marker protein CK19, although tuft cell markers were not detectable at this stage. Upon stimulation with IL-4/13, tuft cell differentiation was confirmed by immunohistochemistry and transcriptomic analysis. We observed induction of DCLK1, as well as taste signaling molecules such as TRPM5 and PLCβ2, which are markers of type II taste cells. Additionally, upregulation of LYZ and DEFB1 mRNA indicated the expression of antimicrobial peptide markers, alongside molecules associated with inflammation. Furthermore, the differentiated organoids specifically responded to a bitter compound, suggesting that pancreatic tuft cells may play a role in detecting potentially harmful chemicals. Finally, immunohistochemical analysis identified tuft cells in the non-human primate pancreas, supporting their involvement in sensing harmful compounds and regulating protective responses within the pancreas.https://www.frontiersin.org/articles/10.3389/fcell.2025.1593226/fulltuft cellsprimateorganoidpancreastype 2 immunity
spellingShingle Kosuke Sakaguchi
Chiemi Kimura-Nakajima
Akihiko Inaba
Yoshiko Hatano
Hanako Ogawa
Yuichi Koshiishi
Keisuke Tanaka
Tatsuya Kometani
Makoto Ohmoto
Koji Sato
Hiroo Imai
Ken Iwatsuki
Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
Frontiers in Cell and Developmental Biology
tuft cells
primate
organoid
pancreas
type 2 immunity
title Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
title_full Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
title_fullStr Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
title_full_unstemmed Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
title_short Generation and functional characterization of tuft cells in non-human primate pancreatic ducts through organoid culture systems
title_sort generation and functional characterization of tuft cells in non human primate pancreatic ducts through organoid culture systems
topic tuft cells
primate
organoid
pancreas
type 2 immunity
url https://www.frontiersin.org/articles/10.3389/fcell.2025.1593226/full
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