A method to differentiate chicken monocytes into macrophages with proinflammatory properties
Lianci Peng1, Robin H G A van den Biggelaar1, Christine A Jansen1
1Department of Biomolecular Health Sciences, Division Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Immunobiology
|November 1, 2020
Summary
This study establishes a 3-day protocol for differentiating chicken monocytes into pro-inflammatory M1-like macrophages. This optimized method aids research into chicken immunity and macrophage polarization.
Area of Science:
- Immunology
- Cell Biology
- Avian Science
Background:
- Macrophages are crucial for innate immunity in pathogen defense.
- Mammalian macrophage differentiation protocols (M1/M2) are established, but chicken protocols lack standardization.
- Understanding chicken macrophage polarization is vital for avian disease research.
Purpose of the Study:
- To develop and characterize a uniform protocol for generating pro-inflammatory M1-like macrophages from chicken blood monocytes.
- To systematically evaluate differentiation over 4 days using morphology, surface markers, and cytokine responses.
- To provide a reliable method for studying chicken macrophage phenotypes.
Main Methods:
- Chicken blood monocytes were cultured for 4 days using granulocyte-macrophage colony-stimulating factor (GM-CSF).
- Daily characterization included cell morphology, surface marker expression (MRC1L-B, MHC-II), and response to Toll-like receptor (TLR) stimulation.
- Functional assays involved lipopolysaccharide (LPS) stimulation for cytokine (IL-1β, IL-6, IL-8) and nitric oxide production, and phagocytosis of E. coli.
Main Results:
- A homogenous, flat, 'fried-egg' morphology was observed on days 3 and 4, similar to human M1 macrophages.
- High expression of MRC1L-B (KUL01) and MHC-II was detected on day 3 and 4 cells.
- LPS stimulation induced pro-inflammatory cytokine gene expression after 3 days, and day 3 macrophages showed phagocytic activity and nitric oxide production.
Conclusions:
- A 3-day culture period is optimal for obtaining pro-inflammatory M1-like macrophages from chicken blood monocytes.
- This standardized protocol enhances understanding of chicken macrophage polarization.
- The findings offer valuable insights for future research on chicken macrophage functions and disease models.
