Liver extracellular matrix hydrogel-based three-dimensional culture system of HepG2 cells to enhance cancer stem cell properties

Both extracellular matrix (ECM) components and three-dimensional (3D) structure play important roles in the expression and maintenance of cancer stem cell (CSC) properties. Considering the excellent biophysical and biochemical properties of hydrogels, the objective of this study was to develop a 3D cell culture system based on liver ECM hydrogel (LEMH) to enhance CSC properties. Results showed that LEMH was devoid of cellular materials but contained the main components of the liver ECM. HepG2 hepatocellular carcinoma cells cultured in LEMH displayed cluster growth and formed multilayer 3D cell structures with increased expression of hepatocyte-specific genes compared to two-dimensional (2D) cells. In addition, enhanced CSC characteristics, including migration, self-renewal and drug-resistance, were observed in 3D cells. More importantly, inhibitory effects of epigallocatechin gallate on CSC self-renewal and metastatic characteristics were observed, confirming the applicability of the LEMH-based 3D model for the research and development of CSC-specific drugs. These findings suggest that LEMH-based 3D culture offers a simple and efficient platform to enhance CSC properties in vitro, thereby providing a novel approach for exploring CSC-specific agents and chemotherapeutic drugs.