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A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma
Published on: September 30, 2021
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The comet assay applied to HepG2 liver spheroids.
Elisabeth Elje1, Michelle Hesler2, Elise Rundén-Pran1
1NILU - Norwegian Institute for Air Research, Department for Environmental Chemistry, Health Effects Laboratory, Instituttveien 18, 2007 Kjeller, Norway.
Mutation Research. Genetic Toxicology and Environmental Mutagenesis
|September 29, 2019
Summary
Advanced 3D cell cultures show promise for predicting chemical toxicity. This study demonstrates 3D HepG2 spheroids are effective for genotoxicity testing, offering a better alternative to 2D models.
Area of Science:
- Toxicology
- Cell Biology
- Biotechnology
Background:
- Reducing in vivo testing is crucial for ethical and efficient chemical safety assessment.
- Advanced in vitro models, particularly 3D cultures, are needed to better predict human toxicity.
- HepG2 cells offer a relevant model for liver toxicity studies.
Purpose of the Study:
- To compare the sensitivity of 2D and 3D HepG2 cell cultures in predicting chemical toxicity.
- To evaluate 3D spheroidal cultures as an advanced in vitro model for genotoxicity testing.
- To assess the utility of the comet assay in 3D hepatocellular cultures.
Main Methods:
- HepG2 spheroids were generated using hanging drop technology.
- Both 2D monolayer and 3D spheroid cultures were exposed to cytotoxic and genotoxic chemicals.
- Cytotoxicity was assessed using alamarBlue assay, flow cytometry, and confocal imaging.
- Genotoxicity was evaluated using the comet assay, with and without Fpg enzyme.
Main Results:
- Cytotoxic and genotoxic responses differed between 2D and 3D HepG2 models.
- 3D spheroidal cultures demonstrated varying sensitivities compared to 2D cultures.
- The comet assay was successfully applied to 3D spheroidal cultures for genotoxicity assessment.
Conclusions:
- Human 3D spheroidal hepatocellular cultures are suitable for genotoxicity testing.
- 3D HepG2 spheroid models represent a promising advancement for in vitro toxicity prediction.
- These models offer a more predictive alternative to traditional 2D cell culture methods.
