Speaker
Description
Introduction:
Accurate preclinical drug evaluation relies on physiologically relevant models that replicate the native tissue microenvironment, including structural architecture and vasculature. Liver cancer, a major global health burden, often requires complex and individualized treatment strategies. However, current therapeutic approaches are typically generalized, failing to address patient-specific responses, which contributes to suboptimal outcomes. Developing reliable liver cancer models that mimic the biological environment is therefore crucial. In particular, the liver’s highly vascularized and metabolically active nature necessitates specialized modeling to capture its complex microenvironment. Traditional 2D cultures and Matrigel-based models fall short in replicating these complexities. Recent advances in 3D bioprinting enable uniform fabrication of multicellular constructs with spatial control, allowing for the integration of functional tissue and vasculature. This study demonstrates the feasibility of using liver decellularized extracellular matrix (dECM) to recreate the tumor microenvironment in hepatocellular carcinoma models. Building on this, we present a 3D bioprinted spheroid platform incorporating tumor cells and endothelial cells, designed for patient-specific drug screening in liver cancer.
Methods:
Liver dECM was used as a bioink to fabricate adherent cells or spheroids containing iPSC-derived hepatocytes, HepG2 cells, or patient-derived liver cancer cells. To evaluate the feasibility of dECM, iPSC-derived hepatocytes were cultured on dECM and Matrigel in both 2D and 3D environments. Albumin and CYP gene expression levels were quantified. For drug screening, HepG2 cells were cultured in 2D with dECM and treated with a panel of 10 anticancer agents. Subsequently, HepG2 and patient-derived tumor cells were encapsulated in liver dECM and bioprinted into 3D spheroids for evaluation with three selected drugs. To assess the effect of vascular proximity, endothelial cells were printed at varying distances from the spheroids, and angiogenic behavior was analyzed.
Results:
In both 2D and 3D conditions, iPSC-derived hepatocytes cultured with liver dECM showed elevated albumin expression compared to those in Matrigel. Notably, CYP gene expression was significantly increased under 3D culture conditions with dECM, indicating enhanced hepatic metabolic function. In case of 2D cultured HepG2, several anticancer drugs showed heightened sensitivity in the presence of dECM. Drug testing in 3D spheroids using HepG2 and patient-derived tumor cells revealed different responses, suggesting improved predictive relevance. Endothelial cells exhibited distance-dependent angiogenic behavior when printed near spheroids, highlighting the spatial influence on vascular growth.
Discussion:
Liver dECM supports hepatic functionality more effectively than Matrigel, especially under 3D conditions that promote metabolic gene expression. The use of 3D bioprinting enabled precise spatial control in fabricating uniform spheroids and integrating vascular components. The observed distance-dependent angiogenesis emphasizes the importance of vascular organization in shaping the tumor microenvironment. Furthermore, modulating spheroid size could induce hypoxic conditions, potentially enhancing angiogenic signaling and promoting the formation of more physiologically relevant vasculature. Because we observed distinct inter-patient variability in drug responses, this integrated platform offers a promising strategy for personalized drug evaluation in liver disease and oncology.
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