Establishment of Melanoma Tumor Xenograft Using Single Cell Line Suspension and Co-injection of Patient-Derived T Cells in Immune-Deficient NSG Mice

  • Fríða Björk GunnarsdóttirEmail author
  • Rolf Kiessling
  • Yago Pico de Coaña
Part of the Methods in Molecular Biology book series (MIMB, volume 1913)


When primary tumor cells are grown in vitro, they are exposed to an environment that is vastly different from the tumor environment they originate from. The in vitro environment can lack the three-dimensional structure of the tumor, other cell types present within the tumor microenvironment, and important growth factors. Humanized mouse models allow researchers to study primary tumor cells in a more natural environment. With further development of several strains of immune-deficient mice, the mouse model allows for observation of the patient-derived tumor xenograft (PDTX) growth alone as well as in the presence of a human immune system. We describe how this can be accomplished with injection of single cell suspension of melanoma tumor cells into immune-deficient NOD-scid IL2Rγnull (NSG) mice. We also describe how tumor cells and immune cells can be co-injected, using Winn assay, and the possibility to use that method to study immune therapies for cancer.

Key words

Xenograft NSG mice Melanoma Immune deficiency Tumor-infiltrating lymphocytes 


  1. 1.
    Morton JJ, Bird G, Refaeli Y, Jimeno A (2016) Humanized mouse xenograft models: narrowing the tumor-microenvironment gap. Cancer Res 76:6153–6158CrossRefGoogle Scholar
  2. 2.
    Schultz LD, Goodwin N, Ishikawa F et al (2014) Human cancer growth and therapy in NOD/SCID/IL2Rγnull (NSG) mice. Cold Spring Harb Protoc 7:694–708Google Scholar
  3. 3.
    Schultz LD, Ishikawa F, Greiner DL (2007) Humanized mice in translational biomedical research. Nat Rev Immunol 7:118–130CrossRefGoogle Scholar
  4. 4.
    Jung J (2014) Human tumor xenograft models for preclinical assessment of anticancer drug development. Toxicol Res 30:1–5CrossRefGoogle Scholar
  5. 5.
    Decker WK, da Silva RD, Sanabria MH et al (2017) Cancer immunotherapy: historical perspective of a clinical revolution and emerging preclinical animal models. Front Immunol 8:829CrossRefGoogle Scholar
  6. 6.
    Simpson-Abelson MR, Sonnenberg GF, Takita H et al (2008) Long-term engraftment and expansion of tumor derived memory T cells following the implantation of non-disrupted pieces of human lung tumor into NOD-scid IL2Rγnull mice. J Immunol 180:7009–7018CrossRefGoogle Scholar
  7. 7.
    Winn HJ (1961) Immune mechanisms in homotransplantation II. Quantitative assay of immunologic activity of lymphoid cells stimulated by tumor homografts. J Immunol 86:228–239PubMedGoogle Scholar
  8. 8.
    Quintana E, Shackleton M, Sabel MS et al (2008) Efficient tumor formation by single human melanoma cells. Nature 456:593–598CrossRefGoogle Scholar
  9. 9.
    Kleinman HK, Martin GR (2005) Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol 15(5):378–386CrossRefGoogle Scholar
  10. 10.
    Fogh J, Fogh JM, Orfeo T (1977) One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J Natl Cancer Inst 59:221–226CrossRefGoogle Scholar
  11. 11.
    Bosma GC, Custer RP, Bosma MJ (1983) A severe combined immunodeficiency mutation in the mouse. Nature 301:527–530CrossRefGoogle Scholar
  12. 12.
    Blunt T, Gell D, Fox M et al (1996) Identification of a nonsense mutation in the carboxyl-terminal region of DNA-dependent protein kinase catalytic subunit in the scid mouse. Proc Natl Acad Sci U S A 93:10285–10290CrossRefGoogle Scholar
  13. 13.
    Schultz LD, Schweitzer PA, Christianson SW et al (1995) Multiple defects in innate and adaptive immunologic function on NOD/LtSz-scid mice. J Immunol 154:180–191Google Scholar
  14. 14.
    Cao X, Shores EW, Hu-Li J et al (1995) Defective lymphoid development in mice lacking expression of the common cytokine receptor gamma chain. Immunity 2:223–238CrossRefGoogle Scholar
  15. 15.
    DiSanto JP, Muller W, Guy-Grand D, Fischer A, Rajewsky K (1995) Lymphoid development in mice with a targeted deletion of the interleukin 2 receptor gamma chain. Proc Natl Acad Sci U S A 92:377–381CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Fríða Björk Gunnarsdóttir
    • 1
    • 2
    Email author
  • Rolf Kiessling
    • 2
  • Yago Pico de Coaña
    • 2
  1. 1.Department of Translational Medicine, Cancer ImmunologyLund UniversityMalmöSweden
  2. 2.Department of Oncology-PathologyKarolinska InstituteStockholmSweden

Personalised recommendations