Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation

Pancreatic ductal adenocarcinoma (PDAC) presents significant treatment challenges due to its desmoplastic reaction, which impedes therapeutic effectiveness, highlighting the need for advanced vitro models to better mimic the complex tumor environment. The current three-dimensional co-culture models...

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Main Authors: Ruben Verloy, Angela Privat-Maldonado, Jonas Van Audenaerde, Sophie Rovers, Hannah Zaryouh, Jorrit De Waele, Delphine Quatannens, Dieter Peeters, Geert Roeyen, Christophe Deben, Evelien Smits, Annemie Bogaerts
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/14/6/450
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author Ruben Verloy
Angela Privat-Maldonado
Jonas Van Audenaerde
Sophie Rovers
Hannah Zaryouh
Jorrit De Waele
Delphine Quatannens
Dieter Peeters
Geert Roeyen
Christophe Deben
Evelien Smits
Annemie Bogaerts
author_facet Ruben Verloy
Angela Privat-Maldonado
Jonas Van Audenaerde
Sophie Rovers
Hannah Zaryouh
Jorrit De Waele
Delphine Quatannens
Dieter Peeters
Geert Roeyen
Christophe Deben
Evelien Smits
Annemie Bogaerts
author_sort Ruben Verloy
collection DOAJ
description Pancreatic ductal adenocarcinoma (PDAC) presents significant treatment challenges due to its desmoplastic reaction, which impedes therapeutic effectiveness, highlighting the need for advanced vitro models to better mimic the complex tumor environment. The current three-dimensional co-culture models of fibroblasts and endothelial cells are lacking, which presents a challenge for performing more comprehensive in vitro research. Our study developed triple co-culture spheroid models using MiaPaCa-2 and BxPC-3 cancer cell lines, with RLT-PSC and hPSC21 pancreatic stellate cell lines and the endothelial cell line HMEC-1. These models were assessed through growth assays, multicolor flow cytometry to optimize cell ratios, cell viability assays to evaluate drug responses, and a tube formation assay with a spheroid-conditioned medium to examine angiogenesis. Our triple co-culture spheroids effectively replicate the PDAC microenvironment, showing significant variations in drug responses influenced by cellular composition, density, and spatial arrangement. The tube formation assay showcased the potential of our models to quantitatively assess a treatment-induced angiogenic response. These cost-effective triple-co-culture in vitro spheroid models provide vital insights into the PDAC microenvironment, significantly improving the quality of the in vitro evaluation of treatment responses.
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issn 2073-4409
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series Cells
spelling doaj-art-d9f6ea6979fb42eb89d28d5655be3ecb2025-08-20T03:43:15ZengMDPI AGCells2073-44092025-03-0114645010.3390/cells14060450Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic EvaluationRuben Verloy0Angela Privat-Maldonado1Jonas Van Audenaerde2Sophie Rovers3Hannah Zaryouh4Jorrit De Waele5Delphine Quatannens6Dieter Peeters7Geert Roeyen8Christophe Deben9Evelien Smits10Annemie Bogaerts11Research Group PLASMANT, Department of Chemistry, University of Antwerp, 2610 Antwerp, BelgiumResearch Group PLASMANT, Department of Chemistry, University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumCenter for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, BelgiumResearch Group PLASMANT, Department of Chemistry, University of Antwerp, 2610 Antwerp, BelgiumPancreatic ductal adenocarcinoma (PDAC) presents significant treatment challenges due to its desmoplastic reaction, which impedes therapeutic effectiveness, highlighting the need for advanced vitro models to better mimic the complex tumor environment. The current three-dimensional co-culture models of fibroblasts and endothelial cells are lacking, which presents a challenge for performing more comprehensive in vitro research. Our study developed triple co-culture spheroid models using MiaPaCa-2 and BxPC-3 cancer cell lines, with RLT-PSC and hPSC21 pancreatic stellate cell lines and the endothelial cell line HMEC-1. These models were assessed through growth assays, multicolor flow cytometry to optimize cell ratios, cell viability assays to evaluate drug responses, and a tube formation assay with a spheroid-conditioned medium to examine angiogenesis. Our triple co-culture spheroids effectively replicate the PDAC microenvironment, showing significant variations in drug responses influenced by cellular composition, density, and spatial arrangement. The tube formation assay showcased the potential of our models to quantitatively assess a treatment-induced angiogenic response. These cost-effective triple-co-culture in vitro spheroid models provide vital insights into the PDAC microenvironment, significantly improving the quality of the in vitro evaluation of treatment responses.https://www.mdpi.com/2073-4409/14/6/450pancreatic ductal adenocarcinomatriple co-culture spheroidstumor microenvironmentdrug resistanceheterogeneityangiogenesis
spellingShingle Ruben Verloy
Angela Privat-Maldonado
Jonas Van Audenaerde
Sophie Rovers
Hannah Zaryouh
Jorrit De Waele
Delphine Quatannens
Dieter Peeters
Geert Roeyen
Christophe Deben
Evelien Smits
Annemie Bogaerts
Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
Cells
pancreatic ductal adenocarcinoma
triple co-culture spheroids
tumor microenvironment
drug resistance
heterogeneity
angiogenesis
title Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
title_full Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
title_fullStr Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
title_full_unstemmed Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
title_short Capturing the Heterogeneity of the PDAC Tumor Microenvironment: Novel Triple Co-Culture Spheroids for Drug Screening and Angiogenic Evaluation
title_sort capturing the heterogeneity of the pdac tumor microenvironment novel triple co culture spheroids for drug screening and angiogenic evaluation
topic pancreatic ductal adenocarcinoma
triple co-culture spheroids
tumor microenvironment
drug resistance
heterogeneity
angiogenesis
url https://www.mdpi.com/2073-4409/14/6/450
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