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Updated: Dec 31, 2025
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
Boyden chamber-based compartmentalized tumor spheroid culture system to implement localized anticancer drug treatment
Shohei Kaneda, Jiro Kawada1, Marie Shinohara1
1Center for International Research on Integrative Biomedical Systems, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 156-0041, Japan.
Abstract:
In anticancer drug development, it is important to simultaneously evaluate both the effect of drugs on cell proliferation and their ability to penetrate tissues. To realize such an evaluation process, here, we present a compartmentalized tumor spheroid culture system utilizing a thin membrane with a through-hole to conduct localized anticancer treatment of tumor spheroids and monitor spheroid dimensions as an indicator of cell proliferation. The system is based on a commercialized Boyden chamber plate; a through-hole was bored through a porous membrane of the chamber, and the pre-existing 0.4 μm membrane pores were filled with parylene C. A HepG2 spheroid was immobilized onto the through-hole, separating the upper and lower compartments. Fluorescein (to verify the isolation between the compartments) and tirapazamine (TPZ; to treat only the lower part of the spheroid) were added to the upper and lower compartments, respectively. Since the transportation of fluorescein was blocked during treatment, i.e., the upper and lower compartments were isolated, it was confirmed that localized TPZ treatment was successfully conducted using the developed system. The effect of localized TPZ treatment on cell proliferation was estimated by measuring the maximum horizontal cross-sectional areas in the upper and lower parts of the spheroid by microscopic observations. This system can, thus, be used to perform localized anticancer drug treatment of tumor spheroids and evaluate the effect of drugs on cell proliferation.
Insights
Researchers developed a novel tumor spheroid culture system for localized anticancer drug treatment. This system enables simultaneous evaluation of drug effects on cell proliferation and tissue penetration, crucial for drug development.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Drug Development
Background:
- Evaluating anticancer drug efficacy requires assessing both cell proliferation inhibition and tissue penetration.
- Current methods may not efficiently integrate these assessments, hindering drug development.
Purpose of the Study:
- To develop a compartmentalized tumor spheroid culture system for localized anticancer treatment and proliferation monitoring.
- To validate the system's ability to perform localized drug delivery and assess treatment effects.
Main Methods:
- A modified Boyden chamber with a parylene C-sealed through-hole was used to culture HepG2 spheroids.
- Localized treatment was achieved by introducing tirapazamine (TPZ) to the lower compartment while maintaining compartment isolation.
- Cell proliferation was monitored by measuring spheroid dimensions via microscopy.
Main Results:
- The system successfully isolated the upper and lower compartments, confirmed by fluorescein transport blockage.
- Localized tirapazamine treatment of the spheroid's lower portion was successfully executed.
- The system allowed for the evaluation of localized TPZ's effect on cell proliferation in different spheroid regions.
Conclusions:
- The developed compartmentalized tumor spheroid culture system facilitates localized anticancer drug treatment.
- This system enables simultaneous evaluation of drug effects on cell proliferation and tissue penetration.
- It offers a valuable tool for advancing anticancer drug development and discovery.
