Protocol for monitoring phagocytosis of cancer cells by TAM-like macrophages using imaging cytometry

Alok K Mishra; Shahid Banday; Ritesh P Thakare; Sunil K Malonia; Michael R Green

doi:10.1016/j.xpro.2024.103320

Protocol for monitoring phagocytosis of cancer cells by TAM-like macrophages using imaging cytometry

STAR Protoc. 2024 Sep 18;5(4):103320. doi: 10.1016/j.xpro.2024.103320. Online ahead of print.

Authors

Alok K Mishra¹, Shahid Banday², Ritesh P Thakare², Sunil K Malonia³, Michael R Green³

Affiliations

¹ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA. Electronic address: [email protected].
² Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA.
³ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA; Cancer Center, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA.

Abstract

Here, we present a protocol for monitoring phagocytosis by M2-type macrophages using automated counting of phagocytic events with an imaging cytometer. We describe steps for isolating and differentiating peripheral blood mononuclear cell (PBMC)-derived monocytes into M2-like macrophages, preparing cancer cells expressing a green fluorescence marker, labeling with a pH-sensitive dye, and co-culturing with macrophages. We then outline procedures for enumerating phagocytic events using an imaging cytometer. For complete details on the use and execution of this protocol, please refer to Mishra et al.¹.

Keywords: Biotechnology and bioengineering; Cancer; Cell Biology; Cell culture; Immunology; Microscopy.

Published by Elsevier Inc.