A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma

The aggressiveness and lack of well-tolerated widely effective treatments for advanced hepatocellular carcinoma (HCC), the predominant form liver cancer, rationalize its rank as second most common cause cancer-related death. Preclinical models need to be adapted recapitulate human conditions select best therapeutic candidates clinical development aid delivery personalized medicine. Three-dimensional (3D) cellular spheroid show promise an emerging in vitro alternative two-dimensional (2D) monolayer cultures. Here, we describe a 3D tumor model which exploits ability individual cells aggregate when maintained hanging droplets, is more representative vivo environment than standard monolayers. Furthermore, spheroids can produced by combining homotypic or heterotypic cells, reflective heterogeneity vivo, potentially enabling study environmental interactions that influence progression treatment responses. current research optimized cell density immobilizing suspensions on lids 10 cm3 Petri dishes. Longitudinal analysis was performed generate growth curves versus tumor/fibroblasts spheroids. Finally, proliferative impact fibroblasts (COS7 cells) myofibroblasts (LX2) (Hep3B) investigated. A seeding 3,000 (in 20 µL media) successfully yielded Huh7/COS7 spheroids, displayed steady increase size up culture day 8, followed retardation. This finding corroborated using Hep3B cultured LX2 (human hepatic stellate line) conditioned medium (CM). CM triggered proliferation compared control In conclusion, this protocol has shown used simple, economical, prescreen tool tumor-stromal comprehensively.