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3D Cell Culture Models as a Platform for Studying Tumor Progression, Testing Treatment Responses, and Discovering Biomarkers.

Peyda Korhan·Ezgi Bağırsakçı·Yasemin Öztemur Islakoğlu·Neşe Atabey
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Abstract

In this chapter, we present a detailed protocol for establishing a three-dimensional (3D) multicellular tumor spheroids (MCTSs) model to simulate the tumor microenvironment (ME) associated with metabolic dysfunction-associated steatotic liver disease (MASLD) for the study of hepatocellular carcinoma (HCC) and colorectal cancer (CRC) cell aggressiveness, growth, and metastasis potential. The MASLD microenvironment (MASLD-ME) is recreated by embedding hepatic stellate cells in a collagen I matrix within a Boyden chamber system. The metabolic medium mimics MASLD conditions, enriched with high glucose, fructose, insulin, and fatty acids, to simulate metabolic stresses associated with the disease.In the protocol, cancer cells are loaded in the upper compartment to analyze their migration toward the MASLD-ME, thereby facilitating studies on cancer cell invasiveness and metastatic capacity. This method offers an adaptable, reproducible model to research disease progression and investigate therapeutic interventions, contributing to preclinical research on MASLD-related liver cancer pathophysiology and potential drug responses.

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HumansSpheroids, CellularTumor MicroenvironmentLiver NeoplasmsDisease ProgressionCarcinoma, HepatocellularCell Culture Techniques, Three DimensionalCell Line, TumorBiomarkers, TumorCell Culture TechniquesColorectal NeoplasmsCell MovementHepatic Stellate CellsColorectal carcinomaHepatocellular carcinomaIn vitro three-dimensional disease modelsMetabolic dysfunction-associated steatohepatitisMetabolic dysfunction-associated steatotic liver diseaseMulticellular tumor spheroidsTumor microenvironmentJournal Article

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Peyda Korhan, Ezgi Bağırsakçı, Yasemin Öztemur Islakoğlu, Neşe Atabey. 2025. 3D Cell Culture Models as a Platform for Studying Tumor Progression, Testing Treatment Responses, and Discovering Biomarkers.. https://doi.org/10.1007/7651_2024_595

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