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Updated: Mar 13, 2026
A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma
Published on: September 30, 2021
Characterization of a functional C3A liver spheroid model
Harriet Gaskell1, Parveen Sharma2, Helen E Colley3
1MRC Centre for Drug Safety Science , Department of Molecular and Clinical Pharmacology , Sherrington Building , Ashton Street and University of Liverpool , L69 3GE , UK . Email: parveen.sharma@liverpool.ac.uk; AstraZeneca , 310 , Cambridge Science Park , Milton Road , Cambridge , Cambridgeshire , CB4 0FZ , UK.
We developed a 3D liver spheroid model using C3A hepatoma cells that shows improved in vivo-like structure and function. This advanced model offers enhanced liver-specific functionality and a more sensitive toxicological response for preclinical drug screening.
Area of Science:
- Hepatology and Toxicology
- Biotechnology and Bioengineering
- Drug Discovery and Development
Background:
- Predictive in vitro liver models are crucial for preclinical screening of hepatotoxic compounds.
- While 3D liver spheroids offer longer functional lifespans than 2D cultures, detailed characterization of their spatiotemporal function and structure is needed for industrial adoption.
Purpose of the Study:
- To develop and characterize a 3D liver spheroid model using C3A hepatoma cells.
- To assess the structural integrity, cellular polarization, zonation features, and functional liver-specific activities of these spheroids.
- To evaluate the spheroid model's response to hepatotoxins compared to traditional 2D cultures.
Main Methods:
- Formation and long-term culture of 3D liver spheroids from C3A hepatoma cells.
- Assessment of spheroid viability, structure, and cellular polarization using microscopy and transporter function assays (e.g., CMFDA substrate transport).
- Measurement of liver-specific functions (albumin and urea synthesis/secretion) and drug penetration (doxorubicin).
- Hepatotoxicity testing with acetaminophen to determine IC50 values in spheroids versus 2D cultures.
Main Results:
- C3A liver spheroids maintained a compact, in vivo-like structure with zonation features for up to 32 days.
- Polarized expression and functional activity of MRP2 and Pgp transporters were observed.
- Spheroids exhibited enhanced synthesis and secretion of albumin and urea, and expressed CYP2E1.
- The spheroid model demonstrated increased susceptibility to hepatotoxins, with a significantly lower IC50 for acetaminophen compared to 2D cultures.
Conclusions:
- A novel method for creating C3A liver spheroids was established, demonstrating cellular polarization and zonation.
- The developed 3D liver spheroid model exhibits superior liver-specific functionality compared to standard 2D models.
- This in vivo-like spheroid model provides a more sensitive toxicological response, making it a valuable tool for preclinical drug screening.
