J Cancer 2011; 2:507-514. doi:10.7150/jca.2.507

Technical Report

Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy

Yen T. Phung1, Dario Barbone2, V. Courtney Broaddus2, Mitchell Ho1 ✉

1. Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America;
2. Lung Biology Center, University of California San Francisco, San Francisco, California, United States of America


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.

Keywords: multicellular spheroids, protocol, monoclonal antibody therapy

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How to cite this article:
Phung YT, Barbone D, Broaddus VC, Ho M. Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy. J Cancer 2011; 2:507-514. doi:10.7150/jca.2.507. Available from http://www.jcancer.org/v02p0507.htm