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

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


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 



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.


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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

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