Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells

Jiajia Tang; Quan Zheng; Qi Wang; Yaru Zhao; Preeta Ananthanarayanan; Chiara Reina; Berina Šabanović; Ke Jiang; Ming-Hsin Yang; Clara Csilla Meny; Huimin Wang; Mette Ø Agerbaek; Thomas Mandel Clausen; Tobias Gustavsson; Chenlei Wen; Felice Borghi; Alfredo Mellano; Elisabetta Fenocchio; Vanesa Gregorc; Anna Sapino; Thor G Theander; Da Fu; Alexandra Aicher; Ali Salanti; Baiyong Shen; Christopher Heeschen

doi:10.1016/j.xpro.2025.103635

Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells

Jiajia Tang, Quan Zheng, Qi Wang, Yaru Zhao, Preeta Ananthanarayanan, Chiara Reina, Berina Šabanović, Ke Jiang, Ming-Hsin Yang, Clara Csilla Meny, Huimin Wang, Mette Ø Agerbaek, Thomas Mandel Clausen, Tobias Gustavsson, Chenlei Wen, Felice Borghi, Alfredo Mellano, Elisabetta Fenocchio, Vanesa Gregorc, Anna SapinoThor G Theander, Da Fu, Alexandra Aicher, Ali Salanti, Baiyong Shen, Christopher Heeschen

Research output: Contribution to journal › Journal article › Research › peer-review

Abstract

We introduce a protocol for generating 3D organoids from circulating tumor cells (CTCs), enabling longitudinal functional and molecular analyses in pancreatic cancer patients, including those with unresectable disease, which constitutes the majority of cases. We outline the process for isolating and characterizing CTCs from the blood of pancreatic cancer patients and provide detailed instructions for initiating, passaging, and phenotyping CTC-derived organoids. Additionally, we describe techniques for utilizing these organoids in drug screening with a focus on stemness-related pathways. For complete details on the use and execution of this protocol, please refer to Tang et al.1.

Original language	English
Article number	103635
Journal	STAR Protocols
Volume	6
Issue number	1
Number of pages	16
ISSN	2666-1667
DOIs	https://doi.org/10.1016/j.xpro.2025.103635
Publication status	Published - 2025

Bibliographical note

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10.1016/j.xpro.2025.103635Licence: CC BY-NC-ND

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Tang, J., Zheng, Q., Wang, Q., Zhao, Y., Ananthanarayanan, P., Reina, C., Šabanović, B., Jiang, K., Yang, M.-H., Meny, C. C., Wang, H., Agerbaek, M. Ø., Clausen, T. M., Gustavsson, T., Wen, C., Borghi, F., Mellano, A., Fenocchio, E., Gregorc, V., ... Heeschen, C. (2025). Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells. STAR Protocols, 6(1), Article 103635. https://doi.org/10.1016/j.xpro.2025.103635

Tang, J, Zheng, Q, Wang, Q, Zhao, Y, Ananthanarayanan, P, Reina, C, Šabanović, B, Jiang, K, Yang, M-H, Meny, CC, Wang, H, Agerbaek, MØ , Clausen, TM , Gustavsson, T, Wen, C, Borghi, F, Mellano, A, Fenocchio, E, Gregorc, V, Sapino, A, Theander, TG, Fu, D, Aicher, A, Salanti, A, Shen, B & Heeschen, C 2025, 'Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells', STAR Protocols, vol. 6, no. 1, 103635. https://doi.org/10.1016/j.xpro.2025.103635

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title = "Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells",

abstract = "We introduce a protocol for generating 3D organoids from circulating tumor cells (CTCs), enabling longitudinal functional and molecular analyses in pancreatic cancer patients, including those with unresectable disease, which constitutes the majority of cases. We outline the process for isolating and characterizing CTCs from the blood of pancreatic cancer patients and provide detailed instructions for initiating, passaging, and phenotyping CTC-derived organoids. Additionally, we describe techniques for utilizing these organoids in drug screening with a focus on stemness-related pathways. For complete details on the use and execution of this protocol, please refer to Tang et al.1.",

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T1 - Protocol for the creation and utilization of 3D pancreatic cancer models from circulating tumor cells

AU - Tang, Jiajia

AU - Zheng, Quan

AU - Wang, Qi

AU - Zhao, Yaru

AU - Ananthanarayanan, Preeta

AU - Reina, Chiara

AU - Šabanović, Berina

AU - Jiang, Ke

AU - Yang, Ming-Hsin

AU - Meny, Clara Csilla

AU - Wang, Huimin

AU - Agerbaek, Mette Ø

AU - Clausen, Thomas Mandel

AU - Gustavsson, Tobias

AU - Wen, Chenlei

AU - Borghi, Felice

AU - Mellano, Alfredo

AU - Fenocchio, Elisabetta

AU - Gregorc, Vanesa

AU - Sapino, Anna

AU - Theander, Thor G

AU - Fu, Da

AU - Aicher, Alexandra

AU - Salanti, Ali

AU - Shen, Baiyong

AU - Heeschen, Christopher

PY - 2025

Y1 - 2025

N2 - We introduce a protocol for generating 3D organoids from circulating tumor cells (CTCs), enabling longitudinal functional and molecular analyses in pancreatic cancer patients, including those with unresectable disease, which constitutes the majority of cases. We outline the process for isolating and characterizing CTCs from the blood of pancreatic cancer patients and provide detailed instructions for initiating, passaging, and phenotyping CTC-derived organoids. Additionally, we describe techniques for utilizing these organoids in drug screening with a focus on stemness-related pathways. For complete details on the use and execution of this protocol, please refer to Tang et al.1.

AB - We introduce a protocol for generating 3D organoids from circulating tumor cells (CTCs), enabling longitudinal functional and molecular analyses in pancreatic cancer patients, including those with unresectable disease, which constitutes the majority of cases. We outline the process for isolating and characterizing CTCs from the blood of pancreatic cancer patients and provide detailed instructions for initiating, passaging, and phenotyping CTC-derived organoids. Additionally, we describe techniques for utilizing these organoids in drug screening with a focus on stemness-related pathways. For complete details on the use and execution of this protocol, please refer to Tang et al.1.

U2 - 10.1016/j.xpro.2025.103635

DO - 10.1016/j.xpro.2025.103635

M3 - Journal article

C2 - 39946239

SN - 2666-1667

VL - 6

JO - STAR Protocols

JF - STAR Protocols

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