Joint infiltrating cell isolation after the induction of CIA
Ankle joints were collected after CIA induction. The skin was removed from the ankle joints and shaken in 5 mL- RPMI 1640 medium (Sigma-Aldrich) containing 10% FBS, 100 units/mL of penicillin, 100 µg/mL streptomycin, 50 µM 2-mercaptoethanol, and 0.385 mg Liberase. The ankle joints were dismantled and single cells were extracted and suspended in culture medium, and isolated with OptiPrep (60 %v/w iodixanol, Takara Bio) according to the manufacturer’s instructions.