In vitro flow cytometry assay to assess primary human and mouse macrophage phagocytosis of live cells

Samantha Y Liu; Naomi Mulugeta; Stephanie K Dougan; Li Qiang

doi:10.1016/j.xpro.2023.102240

In vitro flow cytometry assay to assess primary human and mouse macrophage phagocytosis of live cells

STAR Protoc. 2023 Apr 18;4(2):102240. doi: 10.1016/j.xpro.2023.102240. Online ahead of print.

Authors

Samantha Y Liu¹, Naomi Mulugeta², Stephanie K Dougan³, Li Qiang⁴

Affiliations

¹ Dana-Farber Cancer Institute, Boston, MA 02215, USA.
² Dana-Farber Cancer Institute, Boston, MA 02215, USA; Harvard College, Cambridge, MA 02138, USA.
³ Dana-Farber Cancer Institute, Boston, MA 02215, USA; Harvard Medical School, Boston, MA 02215, USA. Electronic address: stephanie_dougan@dfci.harvard.edu.
⁴ Dana-Farber Cancer Institute, Boston, MA 02215, USA; Harvard Medical School, Boston, MA 02215, USA. Electronic address: li_qiang@dfci.harvard.edu.

Abstract

Although tumor-associated macrophages are generally immunosuppressive, macrophages may also promote tumor clearance via phagocytosis of live tumor cells. Here, we present a protocol for assessing macrophage engulfment of tumor cells in vitro using flow cytometry. We describe steps for cell preparation, reseeding macrophages, and setting up phagocytosis. We then detail procedures for collecting samples, staining macrophages, and flow cytometry. The protocol is applicable to both mouse bone-marrow-derived macrophages and human monocyte-derived macrophages. For complete details on the use and execution of this protocol, please refer to Roehle et al. (2021).¹.

Keywords: Cancer; Cell Biology; Flow Cytometry/Mass Cytometry; Immunology.

Abstract

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