An optimized protocol to determine the engulfment of cancer cells by phagocytes using flow cytometry and fluorescence microscopy

Gi Hoon Nam, Yeonsun Hong, Yoonjeong Choi, Gi Beom Kim, Yoon Kyoung Kim, Yoosoo Yang, In San Kim

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)


The engulfment of cancer cells by macrophages is an important cellular process in innate cancer immunity. Antitumor immunotherapy that utilizes the enhanced engulfment of cancer cells by phagocytic cells has attracted much attention. Therefore, there is a growing demand for methods of measuring cancer cell phagocytosis. Quantifying the various stages of phagocytosis is invaluable for elucidating cancer-immune responses during this process. Here, we describe two phagocytosis assays, a flow cytometric assay and a fluorescent microscopic assay; the flow cytometric method utilizing CellTracker dye provides a simple, measurable, and highly reproducible functional assay to measure the phagocytosis efficiency of cancer cells by bone marrow-derived macrophages. As an alternative method of evaluating various states of cancer cell phagocytosis, a fluorescent microscopic method that employs a pH-sensitive dye (pHrodo-SE dye)is also described in this paper. Image-based analysis using this labeling approach enables researchers to measure phagocytic indices that indicate the number of cancer cells engulfed by each macrophage. We have highlighted that these assays can be applied to multiple tumor types and used as selection tools for a variety of phagocytosis agonist types. The results of this study may facilitate a better understanding of the interactions between tumor cells and phagocytes, which could lead to the identification of new therapeutic targets against cancer.

Original languageEnglish
Pages (from-to)27-32
Number of pages6
JournalJournal of Immunological Methods
Publication statusPublished - 2019 Jul

Bibliographical note

Funding Information:
This work was supported by grants from the National Research Foundation of Korea (NRF) funded by the Korean government ( 2019R1A2B5B03004360 and 2017R1A3B1023418 ), the KU-KIST Graduate School of Converging Science and Technology Program , and the KIST Institutional Program .

Publisher Copyright:
© 2019 Elsevier B.V.


  • Cancer cell
  • Immunotherapy
  • Macrophage
  • Phagocytosis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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