Real-time quantitative PCR analysis of UPR genes in human normal and osteoarthritic chondrocytes

Human chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from individuals within 12 hours of death (normal) and from patients undergoing total knee arthroplasty (osteoarthritic, OA). 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. Total RNA was extracted from the primary 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 normal and three OA human 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 OA samples were compared to the average of the values from the three normal samples