Nam, Gi-Hoon Hong, Yeonsun Choi, Yoonjeong Kim, Gi Beom Kim, Yoon Kyoung Yang, Yoosoo Kim, In-San 2024-01-19T20:00:16Z 2024-01-19T20:00:16Z 2021-09-02 2019-07 0022-1759 https://pubs.kist.re.kr/handle/201004/119864 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. English ELSEVIER MACROPHAGES An optimized protocol to determine the engulfment of cancer cells by phagocytes using flow cytometry and fluorescence microscopy Article 10.1016/j.jim.2019.04.007 1 JOURNAL OF IMMUNOLOGICAL METHODS, v.470, pp.27 - 32 JOURNAL OF IMMUNOLOGICAL METHODS 470 27 32 scie scopus 000483322000005 2-s2.0-85065037271 Biochemical Research Methods Immunology Biochemistry & Molecular Biology Immunology Article MACROPHAGES Phagocytosis Macrophage Cancer cell Immunotherapy