Protocol for generating a pancreatic cancer organoid associated with heterogeneous tumor microenvironment

Kenta Takeuchi; Shunsuke Tabe; Yuya Yamamoto; Kenta Takahashi; Megumi Matsuo; Yasuharu Ueno; Masayuki Ohtsuka; Soichiro Morinaga; Yohei Miyagi; Tomoyuki Yamaguchi; Naoki Tanimizu; Hideki Taniguchi

STAR Protocols (Mar 2025)

Protocol for generating a pancreatic cancer organoid associated with heterogeneous tumor microenvironment

Kenta Takeuchi,
Shunsuke Tabe,
Yuya Yamamoto,
Kenta Takahashi,
Megumi Matsuo,
Yasuharu Ueno,
Masayuki Ohtsuka,
Soichiro Morinaga,
Yohei Miyagi,
Tomoyuki Yamaguchi,
Naoki Tanimizu,
Hideki Taniguchi

Affiliations

Kenta Takeuchi: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan
Shunsuke Tabe: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan; Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8677, Japan
Yuya Yamamoto: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan; Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8677, Japan
Kenta Takahashi: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan; Graduate School of Frontier Sciences, Computational Biology and Medical Science, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8563, Japan
Megumi Matsuo: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan
Yasuharu Ueno: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan
Masayuki Ohtsuka: Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8677, Japan
Soichiro Morinaga: Department of Gastrointestinal Surgery, Kanagawa Cancer Center, 2-3-2 Nakao, Asahi-ku, Yokohama, Kanagawa 241-8515, Japan
Yohei Miyagi: Molecular Pathology and Genetics Division, Kanagawa Cancer Center Research Institute, 2-3-2 Nakao, Asahi-ku, Yokohama, Kanagawa 241-8515, Japan
Tomoyuki Yamaguchi: School of Life Science, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan
Naoki Tanimizu: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan; Corresponding author
Hideki Taniguchi: Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan; Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan; Department of Gastrointestinal Surgery, Kanagawa Cancer Center, 2-3-2 Nakao, Asahi-ku, Yokohama, Kanagawa 241-8515, Japan

Journal volume & issue: Vol. 6, no. 1
p. 103539

Abstract

Read online

Summary: Pancreatic ductal adenocarcinoma (PDAC) organoids that simulate the tumor microenvironment (TME) are an effective tool to identify how TME affects PDAC malignancy. We present a protocol for generating a fused pancreatic cancer organoid (FPCO) that partly reproduces the TME, including heterogeneous cancer-associated fibroblasts (CAFs), using patient-derived PDAC cells and human-induced pluripotent cell-derived endothelial and mesenchymal cells. We also describe the procedure for analyzing FPCO characteristics. FPCO can provide a platform for establishing a reliable drug screening system.For complete details on the use and execution of this protocol, please refer to Takeuchi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords