Gibco™ DMEM, high glucose

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	Synovial tissues were cut into small pieces and digested with collagenase (Sigma-Aldrich, St Louis, MO) in Dulbecco’s modified Eagle’s medium (DMEM) for 2 h at 37 °C to isolate FLS. The FLS were grown in DMEM containing 10 % FetalClone 1 (Thermo Scientific, USA), 100 U/ml penicillin, and 100 mg/ml streptomycin (Sigma-Aldrich, St Louis, MO) in a humidified incubator at 37 °C under 5 % CO2.

Protocol tips
Synovial tissues were cut into small pieces and digested with collagenase (Sigma-Aldrich, St Louis, MO) in Dulbecco’s modified Eagle’s medium (DMEM) for 2 h at 37 °C to isolate FLS. The FLS were grown in DMEM containing 10 % FetalClone 1 (Thermo Scientific, USA), 100 U/ml penicillin, and 100 mg/ml streptomycin (Sigma-Aldrich, St Louis, MO) in a humidified incubator at 37 °C under 5 % CO2.

Publication protocol

Synovial tissue was obtained from patients with RA or osteoarthrosis during joint synovectomies. All patients provided written informed consent. Synovial tissues were cut into small pieces and digested with collagenase (Sigma-Aldrich, St Louis, MO) in Dulbecco’s modified Eagle’s medium (DMEM) for 2 h at 37 °C to isolate FLS. The FLS were grown in DMEM containing 10 % FetalClone 1 (Thermo Scientific, USA), 100 U/ml penicillin, and 100 mg/ml streptomycin (Sigma-Aldrich, St Louis, MO) in a humidified incubator at 37 °C under 5 % CO2. At confluence, the cells were trypsinised and passaged; all cells were used in experiments between passages 3 and 8.

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