Related Experiment Video
Updated: Apr 3, 2026
Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
Published on: April 26, 2024
Microencapsulated Multicellular Tumor Spheroids as a Tool to Test Novel Anticancer Nanosized Drug Delivery Systems In
Abstract:
In the study, MCF-7 human breast adenocarcinoma cells were used to study cytotoxicity of novel anticancer nanosized formulations, such as docetaxel-loaded nanoemulsion and liposomal formulation of a lipophilic methotrexate (MTX) prodrug. In vitro study of cytotoxicity was carried out in 2 models, namely using 3D in vitro model based on multicellular tumor spheroids (MTS) and 2D monolayer culture. MTS were generated by tumor cell cultivation within alginate-oligochitosan microcapsules. In the case of the monolayer culture, cell viability was found to be 25, 18 and 12% for the samples containing nanoemulsion at concentrations 20, 300 and 1000 nM of docetaxel, respectively, after 48 hs incubation. For MTS these values were higher, namely 33, 23 and 18%, respectively. Cytotoxicity of liposomal MTX prodrug-based formulation with final concentration of 1, 2, 10, 50, 100 and 1000 nM in both models was also studied. MTX liposomal formulation demonstrated lower cytotoxicity on MTS compared to intact MTX. Moreover, MTS were also more resistant to both liposomal formulation and intact MTX than the monolayer culture. Thus, at 1000 nM MTX in the liposomal form, cell viability in MTS was 1.4-fold higher than that in the monolayer culture. MTS could be proposed as a promising tool to test novel anticancer nanosized formulations in vitro.
Insights
Novel nanosized anticancer drugs were tested using 3D multicellular tumor spheroids (MTS) and 2D cultures. MTS showed higher resistance to docetaxel nanoemulsions and liposomal methotrexate prodrugs, suggesting their utility in drug development.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Cancer Research
Background:
- Developing novel nanosized formulations for cancer therapy is crucial.
- Evaluating drug efficacy using in vitro models is essential for preclinical studies.
- Multicellular tumor spheroids (MTS) offer a more physiologically relevant 3D in vitro model compared to traditional 2D cultures.
Purpose of the Study:
- To compare the cytotoxicity of docetaxel-loaded nanoemulsions and liposomal methotrexate (MTX) prodrug formulations in 2D monolayer cultures and 3D MTS models.
- To assess the potential of MTS as an in vitro model for evaluating novel anticancer nanosized formulations.
Main Methods:
- MCF-7 human breast adenocarcinoma cells were cultured in 2D monolayers and 3D MTS (within alginate-oligochitosan microcapsules).
- Cytotoxicity of docetaxel nanoemulsion and liposomal MTX prodrug was evaluated at various concentrations after 48 hours of incubation.
- Cell viability was measured in both 2D and 3D models to determine drug efficacy.
Main Results:
- Docetaxel nanoemulsion showed dose-dependent cytotoxicity in both models, with higher cell viability observed in MTS compared to 2D cultures at equivalent concentrations.
- Liposomal MTX prodrug exhibited lower cytotoxicity in MTS than in 2D cultures.
- MTS demonstrated increased resistance to both liposomal MTX and intact MTX compared to monolayer cultures, with a 1.4-fold higher cell viability at 1000 nM MTX in liposomal form.
Conclusions:
- Multicellular tumor spheroids (MTS) represent a promising in vitro tool for evaluating the efficacy of novel anticancer nanosized formulations.
- The 3D microenvironment of MTS influences drug response, highlighting the need for advanced models beyond 2D cultures in anticancer drug development.
- Nanosized formulations may exhibit different cytotoxicity profiles in 3D tumor models compared to traditional 2D cell cultures.
