Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts

Figure 3

Effects of synovial fluid and TNFα on mRNA expression of proinflammatory cytokines in RA fibroblast-like synovial cells. FLS were incubated in absence (FLS) or presence of TNFα (25 ng/mL) (FLS + TNF) or three different synovial fluid pools (FLS + SF) (1 : 8 dilution in culture medium). After 7 days cells were analysed for IL1β, IL6, IL15, and TNFα mRNA expression by quantitative RT-PCR. Data were analysed according to the comparative Ct method and were normalized by GAPDH expression in each sample. Relative mRNA levels were calculated based on the Ct values, corrected for GAPDH expression, according to the equation: [ΔCt = Ct(TNFα, IL1β, IL6, or IL15) − Ct(GAPDH)]. Data are expressed as mean ± SD. versus FLS; versus FLS + TNFα.