153143 20251017144651.0 doi 10.1063/5.0242490 sideral 143573 ART-2025-143573 eng Hernández-Hatibi, Soraya Quantitative characterization of the 3D self-organization of PDAC tumor spheroids reveals cell type and matrix dependence through advanced microscopy analysis 2025 Access copy available to the general public Unrestricted Pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant tumor-associated stroma composed from pancreatic stellate cells, which play a critical role in tumor progression. Developing accurate in vitro models requires understanding the complex interactions between tumor cells and their microenvironment. In this study, we present a quantitative imaging-based characterization of the three dimensional (3D) self-organization of PDAC tumour spheroids using a microfluidic platform that mimics key aspects of the tumor microenvironment. Our model incorporates collagen type I hydrogels to recreate the extracellular matrix, activated human pancreatic stellate cells (HPSCs), and various tumor cell types. Advanced imaging techniques, including Lattice Lightsheet Microscopy, allowed us to analyze the 3D growth and spatial organization of the spheroids, revealing intricate biomechanical interactions. Our results indicate that alterations in matrix properties—such as stiffness, pore size, and hydraulic permeability—due to variations in collagen concentration significantly influence the growth patterns and organization of PDAC spheroids, depending on tumor subtype and epithelial–mesenchymal phenotype. Higher collagen concentrations promoted larger spheroids in epithelial-like cell lines, while mesenchymal-type cells required increased collagen for self-organization into smaller spheroids. Furthermore, coculture with HPSCs affected spheroid formation distinctly based on each PDAC cell line's genetic and phenotypic traits. HPSCs had opposing effects on epithelial-like cell lines: one cell line exhibited enhanced spheroid growth, while another showed inhibited formation, whereas mesenchymal-like spheroids showed minimal impact. These results provide insights into tumor–stroma interactions, emphasizing the importance of the cell-specific and matrix-dependent factors for advancing our understanding of PDAC progression and informing future therapeutic strategies. info:eu-repo/grantAgreement/ES/AEI/FJC2021-048046-I info:eu-repo/grantAgreement/EC/H2020/101018587/EU/Individual and Collective Migration of the Immune Cellular System/ICoMICS This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 101018587-ICoMICS info:eu-repo/grantAgreement/ES/ISCIII/PI20-00625 info:eu-repo/grantAgreement/ES/ISCIII/PI23-00591 info:eu-repo/grantAgreement/ES/MICINN-AEI-FEDER/PID2021-122409OB-C21 info:eu-repo/grantAgreement/ES/MICINN/PRE2019-090264 info:eu-repo/semantics/openAccess by-nc https://creativecommons.org/licenses/by-nc/4.0/deed.es info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Borau, Carlos Universidad de Zaragoza (orcid)0000-0002-3784-1140 Martínez-Bosch, Neus Navarro, Pilar García-Aznar, José Manuel Universidad de Zaragoza (orcid)0000-0002-9864-7683 Guerrero, Pedro Enrique (orcid)0000-0003-2612-9235 5004 605 Universidad de Zaragoza Dpto. Ingeniería Mecánica Área Mec.Med.Cont. y Teor.Est. 9, 1 (2025), 016116 [20 pp.] APL Bioengineering 2473-2877 11394503 http://zaguan.unizar.es/record/153143/files/texto_completo.pdf Versión publicada 2602581 http://zaguan.unizar.es/record/153143/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:153143 articulos driver 2025-10-17-14:36:38 ARTICLE