Isolation of mouse Kupffer cells for phenotypic and functional studies

Area of Science:

• Immunology
• Cell Biology
• Hepatology

Background:

• Kupffer cells are critical liver-resident macrophages involved in immune surveillance and homeostasis.
• Efficient isolation of Kupffer cells is essential for studying their diverse functions and phenotypes.
• Existing protocols may require optimization for specific downstream applications.

Purpose of the Study:

• To provide detailed and reproducible protocols for the isolation of mouse Kupffer cells.
• To enable phenotypic analysis (flow cytometry, mass cytometry, RNA-sequencing) and functional studies of Kupffer cells.
• To offer adaptable procedures for isolating other hepatic cell populations.

Main Methods:

• Detailed step-by-step protocols for Kupffer cell isolation from mouse liver tissue.
• Inclusion of techniques for downstream applications such as flow cytometry, mass cytometry, and RNA-sequencing.
• Adaptable procedures for isolating other liver-resident cell types.

Main Results:

• Validated protocols for high-purity Kupffer cell isolation.
• Demonstrated utility of isolated Kupffer cells in various phenotypic and functional assays.
• Successful adaptation of protocols for isolating other hepatic cell populations.

Conclusions:

• The presented protocols offer a robust method for Kupffer cell isolation and downstream analysis.
• These methods facilitate research into liver immunity and macrophage biology.
• The adaptability of the protocols broadens their utility in hepatic cell research.