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The cell-line-derived subcutaneous tumor model in preclinical cancer research

Nature Protocols volume 17, pages 2108–2128 (2022)Cite this article

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Abstract

Tumor-bearing experimental animals are essential for preclinical cancer drug development. A broad range of tumor models is available, with the simplest and most widely used involving a tumor of mouse or human origin growing beneath the skin of a mouse: the subcutaneous tumor model. Here, we outline the different types of in vivo tumor model, including some of their advantages and disadvantages and how they fit into the drug-development process. We then describe in more detail the subcutaneous tumor model and key steps needed to establish it in the laboratory, namely: choosing the mouse strain and tumor cells; cell culture, preparation and injection of tumor cells; determining tumor volume; mouse welfare; and an appropriate experimental end point. The protocol leads to subcutaneous tumor growth usually within 1–3 weeks of cell injection and is suitable for those with experience in tissue culture and mouse experimentation.

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Fig. 1: Preclinical cancer drug development.
Fig. 2: Different types of subcutaneous model.
Fig. 3: Noninvasive imaging using bioluminescent tumor cells.
Fig. 4: Setting up and using subcutaneous tumors in mice.
Fig. 5: Converting tumor measurements into tumor volumes.

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    Stephen M. Stribbling & Anderson J. Ryan

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Willoughby, C. E. et al. J. Clin. Invest. 130, 258–271 (2020): https://doi.org/10.1172/JCI127483

Masiero, M. et al. Mol. Cancer Ther. 18, 2030–2042 (2019): https://doi.org/10.1158/1535-7163.MCT-18-1176

Jiang, Y. et al. Mol. Cancer Ther. 20, 1663–1671 (2021): https://doi.org/10.1158/1535-7163.MCT-20-085

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Stribbling, S.M., Ryan, A.J. The cell-line-derived subcutaneous tumor model in preclinical cancer research. Nat Protoc 17, 2108–2128 (2022). https://doi.org/10.1038/s41596-022-00709-3

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