Related Experiment Video
Updated: May 9, 2026
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
PDMS well platform for culturing millimeter-size tumor spheroids
Sithira H Ratnayaka1, Taylor E Hillburn, Omid Forouzan
1Dept. of Biomedical Engineering, Tulane University, New Orleans, LA, 70118.
Abstract:
Multicellular tumor spheroids are widely used as in vitro models for testing of anticancer drugs. The advantage of this approach is that it can predict the outcome of a drug treatment on human cancer cells in their natural three-dimensional environment without putting actual patients at risk. Several methods were utilized in the past to grow submillimeter-size tumor spheroids. However, these small models are not very useful for preclinical studies of tumor ablation where the goal is the complete destruction of tumors that can reach several centimeters in diameter in the human body. Here, we propose a PDMS well method for large tumor spheroid culture. Our experiments with HepG2 hepatic cancer cells show that three-dimensional aggregates of tumor cells with a volume as large as 44 mm(3) can be grown in cylindrical PDMS wells after the initial culture of tumor cells by the hanging drop method. This is a 350 times more than the maximum volume of tumor spheroids formed inside hanging drops (0.125 mm(3)).
Insights
Researchers developed a new method to grow large multicellular tumor spheroids for drug testing. This advancement enables more accurate preclinical studies for tumor ablation therapies, improving cancer drug development.
Area of Science:
- Oncology
- Biotechnology
- Drug Discovery
Background:
- Multicellular tumor spheroids are valuable in vitro models for anticancer drug efficacy testing.
- Existing methods for spheroid culture yield small models, limiting their utility for preclinical studies of tumor ablation.
- Large tumor models are needed to accurately simulate in vivo conditions for complete tumor destruction assessment.
Purpose of the Study:
- To develop an improved method for culturing large multicellular tumor spheroids.
- To enable preclinical evaluation of tumor ablation strategies using scaled-up in vitro models.
Main Methods:
- A polydimethylsiloxane (PDMS) well method was developed for large tumor spheroid culture.
- HepG2 hepatic cancer cells were initially cultured using the hanging drop method.
- Subsequent culture was performed in cylindrical PDMS wells to promote spheroid growth.
Main Results:
- The PDMS well method successfully facilitated the growth of large tumor spheroids.
- Tumor spheroids with volumes up to 44 mm³ were achieved, a 350-fold increase over hanging drop methods.
- This method overcomes the limitations of submillimeter-sized spheroids in preclinical research.
Conclusions:
- The proposed PDMS well method enables the scalable culture of large multicellular tumor spheroids.
- This technique provides a more relevant in vitro model for preclinical assessment of tumor ablation therapies.
- The advancement supports more effective and safer anticancer drug development.
