A pancreatic cancer organoid-macrophage co-culture using starPEG-heparin hydrogel deciphers tumor-immune cell interactions

Macrophages are among the most abundant immune cells in the pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment (TME) and play a key role in regulating the immunosuppressive niche that facilitates tumor growth. Although recent three-dimensional (3D) culture systems using patient-derived materials have advanced our understanding of tumor biology, most models lack key cellular TME components and thus fail to capture tumor-immune cell interactions. To address this gap, we developed an in-vitro 3D co-culture model incorporating PDAC patient-derived organoids (PDOs) and macrophages within a synthetic hydrogel matrix. We optimized culture conditions by tuning medium and matrix conditions to support both cell lineages. Flow cytometry and transcriptomic analyses revealed that initially undifferentiated macrophages adopt an M2-like profile upon exposure to PDAC PDOs in starPEG-heparin hydrogels, mirroring the macrophage phenotypes observed by multiplex immunohistochemistry in the matched primary PDAC tissues. Cytokine secretome profiling revealed PDO-specific differences, indicating distinct underlying macrophage polarization subtypes. Collectively, our starPEG-heparin hydrogel-based 3D co-culture enables hypothesis-driven and physiologically relevant studies of tumor-macrophage interactions and may advance immune-modulatory treatment strategies in patients with PDAC.

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Publication Details

Journal
npj Precision Oncology
Published
2026-09-05
DOI
https://doi.org/10.1038/s41698-026-01678-6
Primary Topic
Cancer Cells and Metastasis
Type
article
Field-Weighted Citation Impact
0.00

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article

A pancreatic cancer organoid-macrophage co-culture using starPEG-heparin hydrogel deciphers tumor-immune cell interactions

Maximilian Fusenig, Daniel E. Stange, Daniela Loessner, Mathieu Pecqueux et al.
npj Precision Oncology
Cancer Cells and Metastasis
article

A pancreatic cancer organoid-macrophage co-culture using starPEG-heparin hydrogel deciphers tumor-immune cell interactions

Maximilian Fusenig, Daniel E. Stange, Daniela Loessner, Mathieu Pecqueux, Heike Polster, Carolin Beer, Jürgen Weitz, Sascha Brückmann, Martin Bornhäuser, Anna R. Poetsch, Manuel Pfeifer, Franziska Baenke, Lena Seifert, Daniel Haak, Beatrix Jahnke, Adrian M. Seifert, Marc Schmitz, Antonia Stammberger, Daniela E. Aust, Verena J. Kast, Julius Thyen, Vivian Mittné, Abdul M. Aftab, Carsten Werner, Stefanie Hübner, Rebecca Prause, Victor A. Sioson, Malin H. Reinhart
article en

Abstract

Macrophages are among the most abundant immune cells in the pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment (TME) and play a key role in regulating the immunosuppressive niche that facilitates tumor growth. Although recent three-dimensional (3D) culture systems using patient-derived materials have advanced our understanding of tumor biology, most models lack key cellular TME components and thus fail to capture tumor-immune cell interactions. To address this gap, we developed an in-vitro 3D co-culture model incorporating PDAC patient-derived organoids (PDOs) and macrophages within a synthetic hydrogel matrix. We optimized culture conditions by tuning medium and matrix conditions to support both cell lineages. Flow cytometry and transcriptomic analyses revealed that initially undifferentiated macrophages adopt an M2-like profile upon exposure to PDAC PDOs in starPEG-heparin hydrogels, mirroring the macrophage phenotypes observed by multiplex immunohistochemistry in the matched primary PDAC tissues. Cytokine secretome profiling revealed PDO-specific differences, indicating distinct underlying macrophage polarization subtypes. Collectively, our starPEG-heparin hydrogel-based 3D co-culture enables hypothesis-driven and physiologically relevant studies of tumor-macrophage interactions and may advance immune-modulatory treatment strategies in patients with PDAC.

npj Precision OncologyVol. 10(1)
German Cancer Research Center (DE), University of Cologne (DE), Heidelberg University (DE), Discovery Institute (US), Helmholtz-Zentrum Dresden-Rossendorf (DE), National Center for Tumor Diseases (DE), Leibniz Institute of Polymer Research (DE), Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (DE), Center for Systems Biology Dresden (DE), University Hospital Carl Gustav Carus (DE), Nationales Centrum für Tumorerkrankungen Dresden (DE), Monash University (AU), Technische Universität Dresden (DE)
Deutsche Forschungsgemeinschaft, Bundesministerium für Bildung und Forschung, Deutsche Krebshilfe
Openalex Percentile: Top 13%
Cancer Cells and Metastasis
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