Korhan Peyda, Bağırsakçı Ezgi, Islakoğlu Yasemin Öztemur, Atabey Neşe
Faculty of Medicine, Department of Medical Biology, Izmir Tinaztepe University, Izmir, Turkey.
Galen Research Center, Izmir Tinaztepe University, Izmir, Turkey.
Methods Mol Biol. 2025;2951:267-277. doi: 10.1007/7651_2024_595.
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.
在本章中，我们展示了一个详细的实验方案，用于建立三维（3D）多细胞肿瘤球体（MCTS）模型，以模拟与代谢功能障碍相关的脂肪性肝病（MASLD）相关的肿瘤微环境（ME），用于研究肝细胞癌（HCC）和结直肠癌（CRC）细胞的侵袭性、生长和转移潜能。通过将肝星状细胞嵌入博伊登室系统中的I型胶原基质中来重建MASLD微环境（MASLD-ME）。代谢培养基模拟MASLD条件，富含高糖、果糖、胰岛素和脂肪酸，以模拟与该疾病相关的代谢应激。在该实验方案中，癌细胞加载在上层隔室中，以分析它们向MASLD-ME的迁移，从而便于研究癌细胞的侵袭性和转移能力。该方法提供了一个可适应、可重复的模型，用于研究疾病进展和调查治疗干预措施，有助于对MASLD相关肝癌病理生理学和潜在药物反应的临床前研究。
