Real-time quantitative PCR analysis of UPR genes in human osteoarthritic chondrocytes specimens treated with tunicamycin compared to the untreated (control) chondrocytes

Human osteoarthritic (OA) chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from three patients undergoing total knee arthroplasty. The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. The chondrocytes from each of the three donors were left untreated (control) and treated with tunicamycin (500ng/ml for 20 hours).Total RNA was extracted from the chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z). qPCR gene expression profiling: chondrocytes from three OA donors were used and treated separately as indicated in the summary. Equal amount total RNA from each donor was used for gene expression analysis. The average of the values obtained from the three treated cells were compared to the average of the three untreated control chondrocytes.