Complex Tumor Spheroid Formation and One-Step Cancer-Associated Fibroblasts Purification from Hepatocellular...

Area of Science:

• Biomaterials Science
• Cancer Research
• In Vitro Modeling

Background:

• In vitro cell models are crucial for drug development and toxicity testing, offering alternatives to animal experiments.
• Complex tumor models, particularly for hepatocellular carcinoma (HCC), are essential for predicting drug efficacy and clinical translation.
• Current 3D culture methods often lack the complexity of native tumor microenvironments.

Purpose of the Study:

• To fabricate and evaluate amorphous silicon dioxide (SiO2) surfaces with defined topographical features for culturing hepatocellular carcinoma (HCC) cells.
• To investigate the potential of these novel surfaces to promote complex 2D/3D tissue formation and mimic tumor microenvironments.
• To assess the utility of specific topographies for isolating cancer-associated fibroblasts (CAFs) from HCC tissues.

Main Methods:

• Fabrication of amorphous SiO2 substrates with varied topographical features (pyramids, octahedrons, fractals) in periodic arrays.
• Culture of primary HCC cells and HCC cell lines on these structured surfaces.
• Analysis of cell morphology, tissue architecture (2D/3D formation), and CAF interactions.
• Evaluation of CAF isolation efficiency using specific topographical designs.

Main Results:

• Pyramidal topographies promoted complex 2D/3D tissue formation with primary HCC cells, forming microtissues with CAFs.
• Octahedral and fractal topographies facilitated the exclusion of tumor cells, enabling efficient, one-step isolation of CAFs directly from patient tumor tissue.
• The inorganic surfaces stimulated complex spheroid formation in cell lines rapidly, without requiring additional biological scaffolds.

Conclusions:

• Amorphous SiO2 surfaces with engineered topographies offer a versatile platform for advanced in vitro HCC modeling.
• These novel substrates can recapitulate key aspects of tumor complexity, including CAF invasion and 3D spheroid formation.
• The developed approach provides a scaffold-free method for generating complex tumor models and isolating specific cell populations, advancing preclinical research.