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Probing Single-Cell Macrophage Polarization and Heterogeneity Using Thermo-Reversible Hydrogels in Droplet-Based
B M Tiemeijer1,2, M W D Sweep1,2, J J F Sleeboom2,3,4
1Laboratory of Immunoengineering, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, Netherlands.
Frontiers in Bioengineering and Biotechnology
|November 1, 2021
Summary
This study introduces a novel droplet microfluidics and hydrogel method for culturing single adherent immune cells. This technique reveals hidden cellular heterogeneity and communication drivers, improving understanding of macrophage polarization.
Area of Science:
- Immunology
- Cell Biology
- Microfluidics
- Biomaterials
Background:
- Human immune cells are inherently heterogeneous.
- Understanding single-cell behavior requires eliminating cell-cell communication.
- Droplet microfluidics enables high-throughput single-cell analysis, but adherent cells need a substrate.
Purpose of the Study:
- To develop a method for culturing and analyzing single adherent immune cells at high throughput.
- To investigate macrophage heterogeneity under different polarization states.
- To assess the impact of cell-cell communication on macrophage polarization.
Main Methods:
- Combined droplet microfluidics with a thermo-reversible polyisocyanide (PIC) hydrogel.
- Cultured and stimulated single human macrophages in M1 (pro-inflammatory) and M2 (anti-inflammatory) conditions.
- Utilized phenotypical and functional analysis for single-cell assessment.
Main Results:
- Co-encapsulating multiple cells enhanced macrophage polarization compared to single cells.
- PIC hydrogel droplets improved single macrophage viability and M2 polarization over suspension culture.
- Identified a subset of persistently M1-polarized macrophages, missed in bulk cultures.
Conclusions:
- The combination of droplet microfluidics and hydrogels is effective for studying adherent cell heterogeneity.
- Cellular communication significantly drives macrophage polarization.
- This approach provides high-throughput, single-cell resolution for investigating complex cell behaviors.
