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Updated: Mar 8, 2026
Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing
Published on: April 23, 2015
Studies of anticancer drug cytotoxicity based on long-term HepG2 spheroid culture in a microfluidic system
Agnieszka Zuchowska1, Karina Kwapiszewska1,2, Michal Chudy1
1Department of Microbioanalytics, Institute of Biotechnology, Warsaw University of Technology, Warsaw, Poland.
Abstract:
Cell-on-a-chip systems have become promising devices to study the effectiveness of new anticancer drugs recently. Several microdevices for liver cancer culture and evaluation of the drug cytotoxicity have been reported. However, there are still no proven reports about high-throughput and simple methods for the evaluation of drug cytotoxicity on liver cancer cells. The paper presents the results of the effects of the anticancer drug (5-fluorouracil, 5-FU) on the HepG2 spheroids as a model of liver cancer. The experiments were based on the long-term 3D spheroid culture in the microfluidic system and monitoring of the effect of 5-FU at two selected concentrations (0.5 mM and 1.0 mM). Our investigations have shown that the initial size of the spheroids has influence on the drug effect. With the increase of the spheroids diameter, the drug resistance (for the two tested 5-FU concentrations) decreases. This phenomenon was observed both through cells metabolism analysis, as well as changes in spheroids sizes. In our research, we have shown that the lower 5-FU (0.5 mM) concentration causes higher decrease in HepG2 spheroids viability. Moreover, due to the microsystem construction, we observe the drug resistance effect (10th day of culture) regardless of the initial size of the created spheroids and the drug concentration.
Insights
This study shows that smaller liver cancer spheroids are less resistant to the anticancer drug 5-fluorouracil (5-FU). Lower concentrations of 5-FU more effectively reduce spheroid viability, highlighting spheroid size as a key factor in drug response.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Drug Development
Background:
- Cell-on-a-chip systems offer promising platforms for evaluating anticancer drug efficacy.
- Existing microdevices for liver cancer studies lack high-throughput and simple methods for drug cytotoxicity assessment.
Purpose of the Study:
- To investigate the effects of 5-fluorouracil (5-FU) on HepG2 liver cancer spheroids using a microfluidic system.
- To evaluate the influence of spheroid size on drug cytotoxicity and resistance.
Main Methods:
- Long-term 3D spheroid culture of HepG2 cells within a microfluidic system.
- Monitoring the impact of 5-FU at 0.5 mM and 1.0 mM concentrations on spheroid metabolism and size.
- Analysis of drug resistance across varying initial spheroid diameters.
Main Results:
- Spheroid size significantly influences drug effect; larger spheroids exhibit decreased drug resistance.
- The lower 5-FU concentration (0.5 mM) resulted in a greater reduction in HepG2 spheroid viability.
- A drug resistance effect was observed by the 10th day, irrespective of initial spheroid size or drug concentration, due to the microsystem's design.
Conclusions:
- Spheroid size is a critical determinant of drug response in liver cancer models.
- Microfluidic systems provide a valuable tool for studying drug cytotoxicity and resistance in 3D cancer cultures.
- The findings suggest potential for optimizing drug screening and personalized treatment strategies for liver cancer.
