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Stem Cells International
Volume 2015, Article ID 235192, 14 pages
http://dx.doi.org/10.1155/2015/235192
Research Article

Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages

1Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea
2PWG Genetics Pte Ltd., 15 Tech Park Crescent, Singapore 638117
3Department of Oral and Maxilla Facial Surgery, School of Medicine, Gyeongsang National University, 501 Jinju-daero, Jinju 660-702, Republic of Korea
4Department of Biomedical Sciences, University of Guelph, Guelph, ON, Canada N1G 2W1
5Research Institute of Life Sciences, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea

Received 29 December 2014; Revised 17 March 2015; Accepted 30 March 2015

Academic Editor: Bernard A. J. Roelen

Copyright © 2015 Won-Jae Lee et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 () 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.