title: Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages.
abstract: The identification of stable reference genes is a prerequisite for ensuring accurate validation of gene expression, yet too little is known about stable reference genes of porcine MSCs. The present study was, therefore, conducted to assess the stability of reference genes in porcine MSCs derived from bone marrow (BMSCs), adipose (AMSCs), and skin (SMSCs) with their in vitro differentiated cells into mesenchymal lineages such as adipocytes, osteocytes, and chondrocytes. Twelve commonly used reference genes were investigated for their threshold cycle (Ct) values by qRT-PCR. The Ct values of candidate reference genes were analyzed by geNorm software to clarify stable expression regardless of experimental conditions. Thus, Pearson's correlation was applied to determine correlation between the three most stable reference genes (NF3) and optimal number of reference genes (NFopt). In assessment of stability of reference gene across experimental conditions by geNorm analysis, undifferentiated MSCs and each differentiated status into mesenchymal lineages showed slightly different results but similar patterns about more or less stable rankings. Furthermore, Pearson's correlation revealed high correlation (r > 0.9) between NF3 and NFopt. Overall, the present study showed that HMBS, YWHAZ, SDHA, and TBP are suitable reference genes for qRT-PCR in porcine MSCs.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea ; PWG Genetics Pte Ltd., 15 Tech Park Crescent, Singapore 638117.
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Lee
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Ryoung-Hoon
authors.initials: RH
authors.lastname: Jeon
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Si-Jung
authors.initials: SJ
authors.lastname: Jang
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Ji-Sung
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Seung-Chan
authors.initials: SC
authors.lastname: Lee
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Raghavendra
authors.initials: R
authors.lastname: Baregundi Subbarao
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Sung-Lim
authors.initials: SL
authors.lastname: Lee
authors.affiliation: Department of Oral and Maxilla Facial Surgery, School of Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-702, Republic of Korea.
authors.forename: Bong-Wook
authors.initials: BW
authors.lastname: Park
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea ; Department of Biomedical Sciences, University of Guelph, Guelph, ON, Canada N1G 2W1.
authors.forename: William Allan
authors.initials: WA
authors.lastname: King
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea ; Research Institute of Life Sciences, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
authors.forename: Gyu-Jin
authors.initials: GJ
authors.lastname: Rho