Characterisation of a New Human Alveolar Macrophage-Like Cell Line (Daisy)

Sadofsky, Laura R; Hayman, Yvette A; Vance, Jesse; Cervantes, Jorge L; Fraser, Simon D; Wilkinson, Holly N.; Williamson, James D; Hart, Simon P; Morice, Alyn H.

Abstract

Purpose: There is currently no true macrophage cell line and in vitro experiments requiring these cells currently require mitogenic stimulation of a macrophage precursor cell line (THP-1) or ex vivo maturation of circulating primary monocytes. In this study, we characterise a human macrophage cell line, derived from THP-1 cells, and compare its phenotype to the THP-1 cells. Methods: THP-1 cells with and without mitogenic stimulation were compared to the newly derived macrophage-like cell line (Daisy) using microscopy, flow cytometry, phagocytosis assays, antigen binding assays and gene microarrays. Results: We show that the cell line grows predominantly in an adherent monolayer. A panel of antibodies were chosen to investigate the cell surface phenotype of these cells using flow cytometry. Daisy cells expressed more CD11c, CD80, CD163, CD169 and CD206, but less CD14 and CD11b compared with mitogen-stimulated THP-1 cells. Unlike stimulated THP-1 cells which were barely able to bind immune complexes, Daisy cells showed large amounts of immune complex binding. Finally, although not statistically significant, the phagocytic ability of Daisy cells was greater than mitogen-stimulated THP-1 cells, suggesting that the cell line is more similar to mature macrophages. Conclusions: The observed phenotype suggests that Daisy cells are a good model of human macrophages with a phenotype similar to human alveolar macrophages.

Citation

Sadofsky, L. R., Hayman, Y. A., Vance, J., Cervantes, J. L., Fraser, S. D., Wilkinson, H. N., …Morice, A. H. (2019). Characterisation of a New Human Alveolar Macrophage-Like Cell Line (Daisy). Lung, 197(6), 687-698. https://doi.org/10.1007/s00408-019-00288-3

• Journal Article Type: Article
• Acceptance Date: Oct 25, 2019
• Online Publication Date: Nov 16, 2019
• Publication Date: 2019-12
• Deposit Date: Nov 12, 2019
• Publicly Available Date: Nov 27, 2019
• Journal: Lung
• Print ISSN: 0341-2040
• Electronic ISSN: 1432-1750
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 197
• Issue: 6
• Pages: 687-698
• DOI: https://doi.org/10.1007/s00408-019-00288-3
• Keywords: Macrophage; alveolar; THP-1; Daisy
• Public URL: https://hull-repository.worktribe.com/output/3135430
• Publisher URL: https://link.springer.com/article/10.1007/s00408-019-00288-3