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Updated: May 31, 2026
Macrophage Differentiation and Polarization into an M2-Like Phenotype using a Human Monocyte-Like THP-1 Leukemia Cell Line
Published on: August 2, 2021
A canine macrophage cell line that can be polarized provides a model for canine macrophage differentiation
Qingkang Lyu1,2, Victor P M G Rutten1,3, Ildiko Van Rhijn1,4
1Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 CL, Utrecht, The Netherlands.
Insights
The 030D cell line effectively models canine macrophages, differentiating into M1 and M2 types. This validated tool aids research into canine macrophage biology and immune responses.
Area of Science:
- Veterinary Immunology
- Cell Biology
- Canine Disease Models
Background:
- Macrophage polarization is crucial for immune responses.
- A well-characterized canine macrophage cell line is needed for research.
- The 030D cell line, derived from canine histiocytic sarcoma, was investigated.
Purpose of the Study:
- To assess the macrophage polarization potential of the 030D cell line.
- To characterize M1 and M2 polarized 030D macrophages.
- To compare 030D-derived macrophages with primary canine monocyte-derived macrophages.
Main Methods:
- 030D cells were stimulated with IFN-γ + LPS (M1) or IL-4 (M2).
- Morphology, immunophenotype, gene expression (qPCR), and phagocytic capacity were analyzed.
- 030D-derived macrophages were compared to primary canine monocyte-derived macrophages (MDMs).
Main Results:
- 030D M1 macrophages showed higher expression of M1 surface markers and pro-inflammatory cytokines.
- 030D M2 macrophages exhibited higher expression of M2 markers (MS4A2, CD206).
- 030D M1 macrophages demonstrated superior phagocytic activity compared to M2 macrophages.
Conclusions:
- The 030D cell line is a validated model for studying canine macrophage polarization.
- This cell line provides an accessible tool for investigating canine immune responses.
- 030D-derived macrophages closely resemble primary canine MDMs in key parameters.
Abstract:
Monocyte-like cell lines are valuable models for studying macrophage biology, yet a well-characterized canine macrophage model has been lacking. The canine histiocytic sarcoma-derived cell line 030D was assessed for its macrophage polarization potential by stimulation with IFN-γ + LPS (M1) or IL-4 (M2), or no additives (M0) and characterized for morphology, immunophenotype, marker gene expression, and phagocytic capacity. 030D M1 macrophages expressed higher levels of the surface markers CD32, CD40, CD80, CD83, CD86 and MHC II than 030D M2 macrophages. qPCR analysis showed higher expression of pro-inflammatory cytokines (IL-6, IL-1β, TNF-α, IL-12p35/40, IL-23p19, COX2, CCL2, and CCR7), anti-inflammatory cytokines TGF-β and IL-10, as well as higher expression of LOX-1, LXN, and arginase-1 by 030D M1 macrophages compared to 030D M2 macrophages. Conversely, established M2 markers MS4A2 and CD206 were significantly higher in 030D M2 macrophages compared to 030D M1 macrophages. Functionally, 030D M1 macrophages displayed higher phagocytic activity than 030D M2 macrophages. Comparison of 030D-derived macrophages with primary canine monocyte-derived macrophages (MDMs) demonstrate that both models, with the exception of TGF-β and IL-10, resemble each other in the studied parameters. Thus, the 030D cell line provides a highly accessible and validated tool for studying canine macrophage biology and immune responses.
