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

doi:10.1016/j.xpro.2024.103539

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

STAR Protoc. 2025 Jan 7;6(1):103539. doi: 10.1016/j.xpro.2024.103539. Online ahead of print.

Authors

Affiliations

¹ Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan.
² 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.
³ 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.
⁴ Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
⁵ Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8677, Japan.
⁶ Department of Gastrointestinal Surgery, Kanagawa Cancer Center, 2-3-2 Nakao, Asahi-ku, Yokohama, Kanagawa 241-8515, Japan.
⁷ Molecular Pathology and Genetics Division, Kanagawa Cancer Center Research Institute, 2-3-2 Nakao, Asahi-ku, Yokohama, Kanagawa 241-8515, Japan.
⁸ School of Life Science, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
⁹ Division of Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan. Electronic address: tanimizu@g.ecc.u-tokyo.ac.jp.
¹⁰ 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.

PMID: 39772386
DOI: 10.1016/j.xpro.2024.103539

Abstract

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.

Keywords: Cancer; Organoids; Stem Cells.