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Objective RT-qPCR is a reliable method for gene expression analysis, but the accuracy of the quantitative data depends on the appropriate selection of reference genes. A Co-culture system consisting of periodontal ligament cells (SV-PDL) and cementoblasts (OCCM-30) to investigate the crosstalk between these two cell lines under orthodontic condition is essential for experimental orthodontic setups in-vitro. Therefore, we aimed to identify a set of reliable reference genes suitable for RT-qPCR studies for prospective co-culture systems of OCCM-30 and SV-PDL cells. Results The results demonstrated that  PPIB ,  GUSB  and  RPLP0  turned out to be the three most stable reference genes for OCCM-30 in the co-culture system, while  PPIB ,  POLR2A  and  RPLP0  have the three highest rankings for SV-PDL cells in the co-culture system. The most stable gene combination were  PPIB  and  POLR2A  in the co-culture system. In conclusion,  PPIB  is overall the most stably expressed reference gene for OCCM-30 or SV-PDL cell line in the system. The combination of  PPIB  and  POLR2A  as reference genes are indicated to be the potential and mandatory to obtain accurate quantification results for normalizing RT-qPCR data in genes of interest expression in these two cell lines co-culture systems.

Selection and validation of reference gene for RT-qPCR studies in co-culture system of mouse cementoblasts and periodontal ligament cells