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Updated: May 28, 2026
Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening
Published on: September 5, 2018
Rapid generation of in vitro multicellular spheroids for the study of monoclonal antibody therapy
Yen T Phung1, Dario Barbone, V Courtney Broaddus
11. Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America;
Abstract:
Tumor microenvironments present significant barriers to penetration by antibodies and immunoconjugates and are difficult to study in vitro. Cells cultured as monolayers typically exhibit less resistance to therapy than those grown in vivo. Therefore, it is important to develop an alternative research model that better represents in vivo tumors. We have developed a protocol to produce multicellular spheroids, a simple and more relevant model of in vivo tumors that allows for further investigations of the microenvironmental effects on drug penetration and tumor cell killing. The protocol is used to produce in vitro three-dimensional tumor spheroids from established human cancer cell lines and primary cancer cells isolated from patients without the use of any extracellular components. To study the ability of tumor-targeting immunoconjugates to penetrate these tumor spheroids in vitro, we have used an immunotoxin targeting mesothelin, a surface protein expressed in malignant mesotheliomas. This method for producing consistent, reproducible 3D spheroids may allow for improved testing of novel monoclonal antibodies and other agents for their ability to penetrate solid tumors for cancer therapy.
Insights
Researchers developed a 3D tumor spheroid model to better study drug penetration in vivo. This multicellular spheroid model improves testing of novel cancer therapies targeting solid tumors.
Area of Science:
- Oncology
- Biotechnology
- Cancer Research
Background:
- Tumor microenvironments impede drug delivery and are challenging to study in vitro.
- Standard monolayer cell cultures do not accurately reflect in vivo therapeutic resistance.
- A more representative in vitro model is needed to study drug penetration and efficacy.
Purpose of the Study:
- To develop a protocol for creating reproducible 3D tumor spheroids.
- To establish a more accurate in vitro model for studying drug penetration in solid tumors.
- To investigate the efficacy of immunoconjugates in a 3D tumor model.
Main Methods:
- Developed a protocol for producing in vitro 3D tumor spheroids from human cancer cell lines and primary patient cells.
- Utilized multicellular spheroids as a model system, avoiding extracellular components.
- Employed an immunotoxin targeting mesothelin to assess immunoconjugate penetration in tumor spheroids.
Main Results:
- Successfully produced consistent and reproducible 3D tumor spheroids.
- Demonstrated the utility of the spheroid model for studying drug penetration.
- Showcased the potential for evaluating immunoconjugates targeting specific surface proteins like mesothelin.
Conclusions:
- The developed 3D tumor spheroid protocol offers a relevant in vitro model for cancer research.
- This model facilitates the investigation of microenvironmental effects on therapeutic agent penetration.
- The protocol may enhance the testing of novel monoclonal antibodies and other agents for solid tumor therapy.
