Pen strep amphotericin

J774 and CC (C20A4 -human chondrocyte cell line) were purchased from Sigma Aldrich, as well as tempol, MTT, LPS, antibiotic–antimycotic solution (Pen/Strep/Amphotericin, Sigma-Aldrich Company Ltd, Milan, Italy) and FBS. L-Glutamine was purchased from Euroclone. All stock solutions were made in nonpyrogenic saline (0.9 % NaCl, Baxter, Milan, Italy). All other chemicals were of the highest commercial grade available.

In this study we used two cell lines, murine macrophage cell line (J774) and human chondrocyte cell line (CC) (C20A4 -Sigma-Aldrich, Milan, Italy). J774 cells were maintained in RPMI-1640 culture medium supplemented with 2 mM L-Glutamine, antibiotic–antimycotic solution (Pen/Strep/Amphotericin, Sigma-Aldrich Company Ltd, Milan, Italy), 10% fetal bovine serum (FBS), at 37 °C and 5% CO₂ in a humidified atmosphere. The cells were maintained in culture until 80% confluence and the medium replaced twice a week. CC were grown in complete CC culture medium and maintained in humidified environment at 37 °C and 5% CO₂/95% air atmosphere. The medium was replaced twice a week and cells were cultured until confluence was reached.