Dulbecco s modified eagle s medium

The human lines colon adenocarcinoma (HT29), colorectal carcinoma (HCT116), human fibroblast (HF), endothelial umbilical vein (E926), and sarcoma (SW872 and SW982, provided by ATCC) were all cultured in DMEM (Dulbecco’s modified Eagle’s medium, Eurobio, Les Ulis, France), supplemented with 10% heat-inactivated FBS (Gibco, Life technologies, Milan, Italy), 1% penicillin/streptomycin and 1% Glutammine (Gibco, Life technologies, Milan, Italy). The myxoid sarcoma lines 402–91 WT [22 (link)] and the resistant counterpart 402–91 ET [23 (link)] were maintained in RPMI medium supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin (Gibco, Life technologies, Milan, Italy). All lines grow at 37 °C in a humidified atmosphere with 5% CO₂.

Synoviocytes were grown from synovial tissue samples obtained from RA patients undergoing joint surgery. The patients with RA fulfilled the American College of Rheumatology criteria for RA [20 (link)]. Each individual signed an informed consent form, and the protocol was approved by the committee for the protection of persons participating in biomedical research of the Hospital of Lyon, in compliance with the Helsinki Declaration. Briefly, synovial tissue was minced in small pieces which were allowed to fix on plastic plates. Those samples were maintained in Dulbecco’s modified Eagle’s medium (Eurobio, Courtaboeuf, France) supplemented with 10 % fetal bovine serum (Life Technologies, part of Thermo Fisher Scientific, Carlsbad, CA, USA), 2 % Penicillin-Streptomycin (Eurobio), 1 % L-glutamine (Eurobio), and 1 % Amphotericin B (Eurobio) until cells grew out of the tissue and colonized the plastic dishes. After cells reached confluence, tissue pieces were removed and cells were trypsinized. Synoviocytes were used between passages 4 and 9. Each independent experiment was performed with a different batch of synoviocytes isolated from different RA donors.

The human HepaRG cells were grown in William’s E medium (Sigma, St Louis, MO, USA) supplemented with 10% fetal bovine serum (Life Technologies, Carlsbad, USA), 2 mM L-glutamine (Eurobio, Les Ulis, France), 5 μg/mL insulin (Sigma), 50 μM hydrocortisone hemisuccinate (Serb, Paris, France), 50 U/mL penicillin and 50 μg/mL streptomycin (Eurobio). HepaRG cells were used after 15 days post-plating. Human skin fibroblasts were obtained from biopsies of non-lesional skin as previously described^{8 (link)}. Fibroblasts were maintained in Dulbecco’s modified Eagle’s medium (Eurobio) supplemented with 10% fetal bovine serum (Life Technologies), 2 mM L-glutamine (Eurobio), 100 U/mL penicillin and 100 μg/mL streptomycin (Eurobio).