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

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.
Kenta Takeuchi 1,  Shunsuke Tabe 2,  Yuya Yamamoto 2,  Kenta Takahashi 3,  Megumi Matsuo 4,  Yasuharu Ueno 1,  Masayuki Ohtsuka 5,  Soichiro Morinaga 6,  Yohei Miyagi 7,  Tomoyuki Yamaguchi 8,  Naoki Tanimizu 9,  Hideki Taniguchi 10
Affiliations

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.

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