P51 | DECELLULARIZATION OF HUMAN LUNG SCAFFOLD FOR TUMOR MICROENVIRONMENT MODELLING STUDY

A. Mileo¹, S. Boccia², E. Del Genio¹, R. Pastore¹, M. De Falco³, I. Belviso⁴ | ¹Dept. of Medicine and Health Sciences, University of Molise, Campobasso, Italy; ²Dept. of Biology, University of Naples Federico II, Naples, Italy; ³Dept. of Biology, University of Naples Federico II, Naples, Italy; National Institute of Biostructures and Biosystems (INBB), Rome, Italy; ⁴Dept. of Wellness, Nutrition and Sport, Telematic University Pegaso, Naples, Italy

Decellularized extracellular matrix (d-ECM) is a promising biological scaffold for in vitro cancer research, due to its ability to replicate the native microenvironment and maintain key biomechanical properties¹. An effective decellularization process must remove all cellular material while preserving the ECM’s structure and composition². Human tissue-derived d-ECM offers relevant biochemical and structural cues, making it valuable for disease modelling³. In this study, we developed an in vitro model to investigate the pulmonary metastatic microenvironment induced by breast cancer. Human lung biopsies from pathological donors were snap-frozen, sectioned at 100 μm, and decellularized using 1% SDS and 1% Triton X-100 for 24 h. To prevent contamination, decellularized scaffolds were treated with an antibiotic-antimycotic solution and sterilized with UV light. H&E staining confirmed successful cell removal while maintaining ECM integrity. The lung-derived d-ECM was reseeded with normal human lung fibroblasts, recreating a native-like microenvironment. Human lung fibroblasts were then stimulated with breast cancer conditioned medium to study how stromal cells remodel the ECM and support tumor invasion. This model offers a reliable platform to explore cell–matrix interactions and the stromal role in cancer progression, providing new insights for preclinical oncology research.