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A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma
Published on: September 30, 2021
Constrained spheroids for prolonged hepatocyte culture.
Wen Hao Tong1, Yu Fang1, Jie Yan2
1Institute of Biotechnology and Nanotechnology, A*STAR, The Nanos, #04-01, 31 Biopolis Way, Singapore 138669, Singapore; NUS Graduate School for Integrative Sciences and Engineering, Centre for Life Sciences (CeLS), #05-01, 28 Medical Drive, Singapore 117456, Singapore; Department of Physiology, Yong Loo Lin School of Medicine, MD9-03-03, 2 Medical Drive, Singapore 117597, Singapore.
A novel Constrained Spheroids (CS) culture system minimizes cell loss in liver spheroids. This improved platform enhances hepatocyte function for more reliable drug screening assays.
Area of Science:
- Hepatology and Cell Culture Technology
- Biomaterials and Tissue Engineering
Background:
- Maintaining liver-specific functions in primary hepatocytes in vitro is crucial for drug screening.
- Hepatocyte spheroid cultures improve function but suffer from cell loss, causing assay variability.
- Existing static and perfusion methods lead to spheroid detachment and cell loss.
Purpose of the Study:
- To develop a novel culture system to prevent cell loss in hepatocyte spheroids.
- To enhance the stability and functionality of in vitro liver models.
- To reduce variability in downstream assays like drug screening.
Main Methods:
- Development of a Constrained Spheroids (CS) culture system immobilizing spheroids between a modified glass coverslip and a porous Parylene C membrane.
- Surface modification with poly(ethylene glycol) and galactose ligands for optimal spheroid formation and maintenance.
- Evaluation of cell loss and hepatocyte functions under static and perfusion conditions compared to standard models.
Main Results:
- The CS culture system significantly minimized cell loss, even under perfusion.
- Hepatocytes cultured as CS spheroids exhibited enhanced urea secretion and metabolic activity (CYP1A1, CYP3A2).
- CS culture demonstrated superior performance compared to the standard collagen sandwich model.
Conclusions:
- The Constrained Spheroids (CS) culture system effectively prevents cell loss in hepatocyte spheroids.
- CS culture enhances key liver-specific functions, offering a more robust platform for in vitro studies.
- This system represents an improvement over current hepatocyte spheroid-based culture methods for drug discovery and research.
