Time-Lapse 2D Imaging of Phagocytic Activity in M1 Macrophage-4T1 Mouse Mammary Carcinoma Cells in Co-cultures
| Autor: | Nur Aima Hafiza Shazali, Mohd Azuraidi Osman, Adam Leow Thean Chor, Noorzaileen Eileena Zaidi, Kamariah Ibrahim, Nik Mohd Afizan Nik Abd Rahman, Marilyn Jaoi-Edward |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
General Chemical Engineering medicine.medical_treatment Breast Neoplasms General Biochemistry Genetics and Molecular Biology Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Immune system Phagocytosis Cancer immunotherapy Live cell imaging Cell Line Tumor Tumor Microenvironment medicine Animals Humans Cytotoxic T cell Macrophage Tumor microenvironment General Immunology and Microbiology Chemistry Macrophages General Neuroscience Carboxyfluorescein succinimidyl ester Coculture Techniques Cell biology 030104 developmental biology 030220 oncology & carcinogenesis Cancer cell Female |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/60281 |
| Popis: | Tumor-associated macrophages (TAMs) have been identified as an important component for tumor growth, invasion, metastasis, and resistance to cancer therapies. However, tumor-associated macrophages can be harmful to the tumor depending on the tumor microenvironment and can reversibly alter their phenotypic characteristics by either antagonizing the cytotoxic activity of immune cells or enhancing anti-tumor response. The molecular actions of macrophages and their interactions with tumor cells (e.g., phagocytosis) have not been extensively studied. Therefore, the interaction between immune cells (M1/M2-subtype TAM) and cancer cells in the tumor microenvironment is now a focus of cancer immunotherapy research. In the present study, a live cell coculture model of induced M1 macrophages and mouse mammary 4T1 carcinoma cells was developed to assess the phagocytic activity of macrophages using a time-lapse video feature using phase-contrast, fluorescent, and differential interference contrast (DIC) microscopy. The present method can observe and document multipoint live-cell imaging of phagocytosis. Phagocytosis of 4T1 cells by M1 macrophages can be observed using fluorescent microscopy before staining 4T1 cells with carboxyfluorescein succinimidyl ester (CFSE). The current publication describes how to coculture macrophages and tumor cells in a single imaging dish, polarize M1 macrophages, and record multipoint events of macrophages engulfing 4T1 cells during 13 h of coculture. |
| Databáze: | OpenAIRE |
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