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Dulbecco's Modified Eagle Medium (Life Technologies; UK) containing 10% foetal bovine serum (FBS), 0.2% amphotericin B (2 µg/ml), and 1% penicillin (100 U/ml) streptomycin (100 µg/ml) at 37°C in 5% CO2. |
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Protocol tips |
Dulbecco's Modified Eagle Medium (Life Technologies; UK) containing 10% foetal bovine serum (FBS), 0.2% amphotericin B (2 µg/ml), and 1% penicillin (100 U/ml) streptomycin (100 µg/ml) at 37°C in 5% CO2. |
Publication protocol
Forelimbs from three horses were collected from a local abattoir. The metacarpophalangeal joint was cleaned and skinned before opening. The joint surface was scored macroscopically to ensure absence of orthopaedic disease.3 Synovium was dissected, finely diced, and cells isolated using 0.25% (w/v) trypsin/0.1% (w/v) EDTA digestion followed by incubation with 2 mg/ml collagenase type 2 (Worthington Biochemicals; Berkshire, UK) Cells were seeded at 6 × 104 cells/cm2 and expanded for up to three passages using split ratios of 1:3. Unless stated otherwise, all cells were cultured in Dulbecco's Modified Eagle Medium (Life Technologies; UK) containing 10% foetal bovine serum (FBS), 0.2% amphotericin B (2 µg/ml), and 1% penicillin (100 U/ml) streptomycin (100 µg/ml) at 37°C in 5% CO2.
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Paper title
Interaction with macrophages attenuates equine fibroblast‐like synoviocyte ADAMTS5 (aggrecanase‐2) gene expression following inflammatory stimulation
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