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