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Updated: Aug 16, 2026
Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
Published on: April 26, 2024
Development of an in vitro multicellular tumor spheroid model using microencapsulation and its application in
Xulang Zhang1, Wei Wang, Weiting Yu
1Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
Abstract:
In this study, an in vitro multicellular tumor spheroid model was developed using microencapsulation, and the feasibility of using the microencapsulated multicellular tumor spheroid (MMTS) to test the effect of chemotherapeutic drugs was investigated. Human MCF-7 breast cancer cells were encapsulated in alginate-poly-l-lysine-alginate (APA) microcapsules, and a single multicellular spheroid 150 mum in diameter was formed in the microcapsule after 5 days of cultivation. The cell morphology, proliferation, and viability of the MMTS were characterized using phase contrast microscopy, BrdU-labeling, MTT stain, calcein AM/ED-2 stain, and H&E stain. It demonstrated that the MMTS was viable and that the proliferating cells were mainly localized to the periphery of the cell spheroid and the apoptotic cells were in the core. The MCF-7 MMTS was treated with mitomycin C (MC) at a concentration of 0.1, 1, or 10 times that of peak plasma concentration (ppc) for up to 72 h. The cytotoxicity was demonstrated clearly by the reduction in cell spheroid size and the decrease in cell viability. The MMTS was further used to screen the anticancer effect of chemotherapeutic drugs, treated with MC, adriamycin (ADM) and 5-fluorouracil (5-FU) at concentrations of 0.1, 1, and 10 ppc for 24, 48, and 72 h. MCF-7 monolayer culture was used as control. Similar to monolayer culture, the cell viability of MMTS was reduced after treatment with anticancer drugs. However, the inhibition rate of cell viability in MMTS was much lower than that in monolayer culture. The MMTS was more resistant to anticancer drugs than monolayer culture. The inhibition rates of cell viability were 68.1%, 45.1%, and 46.8% in MMTS and 95.1%, 86.8%, and 91.6% in monolayer culture treated with MC, ADM, and 5-FU at 10 ppc for 72 h, respectively. MC showed the strongest cytotoxicity in both MMTS and monolayer, followed by 5-FU and ADM. It demonstrated that the MMTS has the potential to be a rapid and valid in vitro model to screen chemotherapeutic drugs with a feature to mimic in vivo three-dimensional (3-D) cell growth pattern.
Insights
This study developed a microencapsulated multicellular tumor spheroid (MMTS) model for drug screening. The MMTS demonstrated resistance to chemotherapy compared to traditional monolayer cultures, offering a more realistic in vitro cancer model.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Drug Discovery
Background:
- Traditional 2D cell cultures do not accurately mimic the in vivo tumor microenvironment.
- Developing advanced in vitro models is crucial for effective drug screening and personalized medicine.
Purpose of the Study:
- To develop and validate a microencapsulated multicellular tumor spheroid (MMTS) model for evaluating chemotherapeutic drug efficacy.
- To compare the drug response of MMTS with conventional MCF-7 breast cancer cell monolayer cultures.
Main Methods:
- Human MCF-7 breast cancer cells were encapsulated in alginate-poly-l-lysine-alginate (APA) microcapsules to form MMTS.
- MMTS morphology, proliferation, and viability were assessed using microscopy and staining techniques.
- MMTS and monolayer cultures were treated with mitomycin C (MC), adriamycin (ADM), and 5-fluorouracil (5-FU) at varying concentrations and time points.
Main Results:
- The MMTS model successfully mimicked in vivo three-dimensional (3-D) cell growth, with proliferating cells at the periphery and apoptotic cells in the core.
- MMTS exhibited significantly lower inhibition rates of cell viability compared to monolayer cultures when treated with chemotherapeutic drugs.
- Mitomycin C demonstrated the strongest cytotoxicity, followed by 5-FU and ADM, in both models, with MMTS showing greater drug resistance.
Conclusions:
- The MMTS model provides a more physiologically relevant in vitro platform for anticancer drug screening.
- This model's ability to mimic 3-D tumor growth offers potential for more accurate prediction of drug response.
- The MMTS model can serve as a rapid and valid tool for screening chemotherapeutic drugs, potentially improving preclinical drug development.
