Protocol for detecting macrophage-mediated cancer cell phagocytosis in vitro and in vivo

Chuan Xu; Hong Wu; Yiqiang Liu; Feifei Li; Rajesh Kumar Manne; Hui-Kuan Lin

doi:10.1016/j.xpro.2022.101940

Protocol for detecting macrophage-mediated cancer cell phagocytosis in vitro and in vivo

STAR Protoc. 2023 Mar 17;4(1):101940. doi: 10.1016/j.xpro.2022.101940. Epub 2022 Dec 14.

Authors

Chuan Xu¹, Hong Wu², Yiqiang Liu², Feifei Li², Rajesh Kumar Manne³, Hui-Kuan Lin⁴

Affiliations

¹ Department of Oncology, Sichuan Academy of Medical Sciences, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 610072, China; Integrative Cancer Center & Cancer Clinical Research Center, Sichuan Cancer Hospital & Institute Sichuan Cancer Center, School of Medicine University of Electronic Science and Technology of China, Chengdu, Sichuan 610047, China; Department of Cancer Biology, Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA. Electronic address: xuchuan100@163.com.
² Department of Oncology, Sichuan Academy of Medical Sciences, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 610072, China; Integrative Cancer Center & Cancer Clinical Research Center, Sichuan Cancer Hospital & Institute Sichuan Cancer Center, School of Medicine University of Electronic Science and Technology of China, Chengdu, Sichuan 610047, China.
³ Department of Cancer Biology, Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA.
⁴ Department of Cancer Biology, Department of Cancer Biology, Wake Forest University School of Medicine, Winston Salem, NC 27157, USA. Electronic address: hulin@wakehealth.edu.

Abstract

Here, we present optimized approaches to identify the efficiency of cancer cell phagocytosis by macrophages in vitro and in vivo. We describe the preparation and co-culture of macrophages and cancer cells, followed by in vitro phagocytosis assay using flow cytometry and confocal microscopy, respectively. We then detail the establishment of xenograft tumor mouse model and the in vivo detecting of phagocytosis efficiency by flow cytometry and qRT-PCR. This protocol provides a convenient way to assess macrophage-mediated phagocytosis of cancer cells. For complete details on the use and execution of this protocol, please refer to Xu et al.¹.

Keywords: Cancer; Cell Biology; Cell culture; Flow Cytometry/Mass Cytometry; Immunology; Microscopy; Model Organisms; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Coculture Techniques
Cytophagocytosis*
Disease Models, Animal
Humans
Macrophages
Mice
Neoplasms*
Phagocytosis