Oxidative Stress Gene Expression in DBMSCs

The expression of 84 genes related to oxidative stress (catalogue number 330231 Qiagen, Hilden, Germany) by DBMSCs was determined using QuantiTect Primer Assay (Qiagen) in a real-time polymerase chain reaction (RT-PCR) as previously published [1 (link)]. Briefly, total RNA from DBMSCs treated with or without H₂O₂ was isolated, and cDNA was then synthesized using FastLane Cell cDNA kit and RT Primer Mix (Qiagen) as previously published [1 (link)]. After quantifying mRNA using QuantiTect SYBR Green PCR kit (Qiagen), the real-time PCR reaction was performed in triplicate on the CFX96 real-time PCR detection system (Bio-Rad) as previously published [1 (link)]. Then, the data were analyzed using the CFX manager software (Bio-Rad). The results were exported to Microsoft Excel for further analysis. The results were expressed in terms of fold change by calculating the ΔΔ⁻² values. The relative expression an internal housekeeping gene as a loading control was used as provided in the kit. Each experiment was carried out using DBMSCs (passage 3) prepared from three individual placentae.