Protocol for preparation of mouse synovium for flow cytometry and RNA-seq

Area of Science:

• Immunology and Rheumatology
• Cell Biology
• Genomics

Background:

• Mouse synovium models rheumatoid arthritis and osteoarthritis.
• Sterile inflammation is a key feature of these joint diseases.

Purpose of the Study:

• To present a detailed protocol for inducing arthritis in mice.
• To outline methods for harvesting and preparing single-cell suspensions from mouse synovium.
• To enable downstream molecular analyses of synovial cells.

Main Methods:

• Induction of arthritis in mouse models.
• Synovium harvesting and dissociation.
• Preparation of single-cell suspensions.
• Intravascular labeling (optional).

Main Results:

• A reproducible protocol for obtaining single-cell suspensions from mouse synovium.
• The protocol supports both steady-state and inflamed conditions.
• The resulting cells are suitable for various high-throughput analyses.

Conclusions:

• This protocol facilitates the study of synovial cell populations in arthritis.
• It enables detailed molecular characterization using techniques like single-cell RNA sequencing.
• Provides a foundation for research into pathogenesis and therapeutic targets in joint diseases.