Advertisement

Tumor-Associated Macrophage Isolation and In Vivo Analysis of Their Tumor-Promoting Activity

  • Nicholas M. Clark
  • Paula D. Bos
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1884)

Abstract

Characterization of individual cell populations from the tumor microenvironment is critical to understand their functional contribution to tumor progression. Magnetic bead enrichment and fluorescence-activated cell sorting (FACS) allow for the isolation of specific cell types that can be used in downstream applications, including in vitro and in vivo functional studies and molecular profiling. In this chapter, we describe the process of isolation of tumor-associated macrophages (TAMs) from primary murine breast tumors subsequent to therapeutic or experimental intervention. Additionally, we further detail how to analyze their ability to support tumor cell growth by co-injecting isolated TAMs with tumor cells orthotopically into the mammary gland of immune-deficient hosts, and monitoring tumor progression by live imaging and caliper measurement.

Key words

Tumor-associated macrophages (TAMs) Magnetic bead enrichment Fluorescence-activated cell sorting (FACS) Orthotopic co-injection Myeloid effector function Live imaging Tumor monitoring 

Notes

Acknowledgments

We thank members of the laboratory for insightful comments on the manuscript. Work in the Bos laboratory is partially supported by grants from METAvivor Inc., American Cancer Society Institutional Research Grants, and the Massey Cancer Center.

References

  1. 1.
    Wynn TA, Chawla A, Pollard JW (2013) Macrophage biology in development, homeostasis and disease. Nature 496:445–455CrossRefGoogle Scholar
  2. 2.
    Noy R, Pollard JW (2014) Tumor-associated macrophages: from mechanisms to therapy. Immunity 41:49–61CrossRefGoogle Scholar
  3. 3.
    Biswas SK, Mantovani A (2010) Macrophage plasticity and interaction with lymphocyte subsets: cancer as a paradigm. Nat Immunol 11:889–896CrossRefGoogle Scholar
  4. 4.
    Ruffell B, Affara NI, Coussens LM (2012) Differential macrophage programming in the tumor microenvironment. Trends Immunol 33:119–126CrossRefGoogle Scholar
  5. 5.
    Ruffell B, Coussens LM (2015) Macrophages and therapeutic resistance in cancer. Cancer Cell 27:462–472CrossRefGoogle Scholar
  6. 6.
    Kocaturk B, Versteeg HH (2015) Orthotopic injection of breast cancer cells into the mammary fat pad of mice to study tumor growth. J Vis ExpGoogle Scholar
  7. 7.
    Denardo DG, Brennan DJ, Rexhepaj E et al (2011) Leukocyte complexity predicts breast cancer survival and functionally regulates response to chemotherapy. Cancer Discov 1:54–67CrossRefGoogle Scholar
  8. 8.
    Liu Y, Chen K, Wang C, Gong W, Yoshimura T, Wang JM, Liu M (2013) Isolation of mouse tumor-infiltrating leukocytes by percoll gradient centrifugation. Bio-protocol 3Google Scholar
  9. 9.
    Minn AJ, Gupta GP, Siegel PM et al (2005) Genes that mediate breast cancer metastasis to lung. Nature 436:518–524CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of PathologyIntegrative Life Sciences Graduate Program, Virginia Commonwealth University School of MedicineRichmondUSA
  2. 2.Department of PathologyMassey Cancer Center, Virginia Commonwealth University School of MedicineRichmondUSA

Personalised recommendations