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Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice

Nature Protocols volume 4, pages 1670–1680 (2009)Cite this article

Abstract

For decades, xenografts using well-established human tumor cell lines have been the most commonly used models to study human cancers in mice. More recently, human tumors implanted directly into immunodeficient mice have become increasingly popular as evidence accrues that they more accurately recapitulate features of patient tumors. Here we describe our protocols for the orthotopic and heterotopic implantation of pancreatic cancer cell lines and freshly isolated patient tumors into immunodeficient mice. We also describe procedures for the digestion of tumors into single-cell suspensions for the isolation of subpopulations of tumor cells. Orthotopic or heterotopic implantation of established cell lines requires 1–2 h, with 1-cm tumors arising after 2–5 weeks. Engraftment of patient tumor samples takes ∼2 h and growth of palpable tumor requires ∼14 weeks. Once established, direct xenograft tumors require 2 and 5 h for heterotopic and orthotopic implantation, respectively, and 5–6 weeks for palpable tumor growth.

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Figure 1: Comparison of current xenograft models.
Figure 2: Histological appearance of direct heterotopic and orthotopic xenograft tumors (H&E).
Figure 3: Heterotopic implantation of tumor.
Figure 4: Single-layer closure of mouse skin with interrupted sutures.
Figure 5: Orthotopic implantation of pancreatic cancer cells.
Figure 6: Orthotopic injection of pancreatic cancer cells.
Figure 7: Two-layer closure of mouse abdominal wall with interrupted sutures.
Figure 8: Time until palpable tumor formation varies with xenograft generation.

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Acknowledgements

This work was supported by the Various Donor Fund for Pancreatic Cancer Research, M.D. Anderson Cancer Center (J.B.F. and D.B.E.), NIH T-32 09599 (M.P.K.), and NIH 5P20A101936 (G.E.G. and J.L.A.).

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Author notes

  1. Jason B Fleming and Gary E Gallick: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Surgical Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    Michael P Kim, Douglas B Evans & Jason B Fleming

  2. Department of Cancer Biology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    Michael P Kim

  3. Department of Pathology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    Huamin Wang

  4. Department of Gastrointestinal Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    James L Abbruzzese

  5. Department of Genitourinary Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA

    Gary E Gallick

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  1. Michael P Kim
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Contributions

M.P.K., J.B.F. and G.E.G. designed experiments and analyzed data; M.P.K. carried out the animal study and conducted experiments; M.P.K., J.L.A. and G.E.G. wrote the paper; D.B.E., J.B.F. and H.W. acquired and processed surgical specimens; and H.W. examined pathologic specimens.

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Correspondence to Gary E Gallick.

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Kim, M., Evans, D., Wang, H. et al. Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice. Nat Protoc 4, 1670–1680 (2009). https://doi.org/10.1038/nprot.2009.171

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