P-73: Effect of Donor Age on The Expression Stability of GAPDH as A ReferenceGene for Gene Expression Analysis ofEquine Adipose-Derived Mesenchymal Stem Cells

Background: Adipose tissue is a main source for isolation of equine mesenchymal stem cells (MSCs) at different ages. It seems that characteristics of adipose-derived MSCs especially gene expression profile are changing along with age increase. A proper reference gene is required for normalizing data in gene expression analysis by qRT-PCR. This study aimed to evaluate whether GAPDH has a stable expression in adipose-derived MSCs isolated from horses with different ages. Materials and Methods: MSCs isolated from adipose tissue of 3 mares, aged 3, 6 and 9 years old. After 3 passages, total RNA of cultured cells was isolated and mRNA reverse transcribed to cDNA. qRT-PCR was done using GAPDH primers in triplicate. Results: Our analysis indicated that Ct of GAPDH gene is very similar in all samples. Thus, its expression was almost the same at adipose-derived MSCs of horses with different ages. Mean ± SD of Ct was 15.53 ± 0.47. Conclusion: Based on the results, it suggests that expression level of GAPDH is stable in MSCs of horses with different ages. Thus, it can be considered as a suitable reference gene for gene expression analysis for adipose-derived MSCs isolated at different ages.