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

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 fro...

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Autores principales: Lee, Won-Jae, Jeon, Ryoung-Hoon, Jang, Si-Jung, Park, Ji-Sung, Lee, Seung-Chan, Baregundi Subbarao, Raghavendra, Lee, Sung-Lim, Park, Bong-Wook, King, William Allan, Rho, Gyu-Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4417979/
https://www.ncbi.nlm.nih.gov/pubmed/25972899
http://dx.doi.org/10.1155/2015/235192
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author Lee, Won-Jae
Jeon, Ryoung-Hoon
Jang, Si-Jung
Park, Ji-Sung
Lee, Seung-Chan
Baregundi Subbarao, Raghavendra
Lee, Sung-Lim
Park, Bong-Wook
King, William Allan
Rho, Gyu-Jin
author_facet Lee, Won-Jae
Jeon, Ryoung-Hoon
Jang, Si-Jung
Park, Ji-Sung
Lee, Seung-Chan
Baregundi Subbarao, Raghavendra
Lee, Sung-Lim
Park, Bong-Wook
King, William Allan
Rho, Gyu-Jin
author_sort Lee, Won-Jae
collection PubMed
description 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 (NF(3)) and optimal number of reference genes (NF(opt)). 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 NF(3) and NF(opt). Overall, the present study showed that HMBS, YWHAZ, SDHA, and TBP are suitable reference genes for qRT-PCR in porcine MSCs.
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spelling pubmed-44179792015-05-13 Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages Lee, Won-Jae Jeon, Ryoung-Hoon Jang, Si-Jung Park, Ji-Sung Lee, Seung-Chan Baregundi Subbarao, Raghavendra Lee, Sung-Lim Park, Bong-Wook King, William Allan Rho, Gyu-Jin Stem Cells Int Research Article 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 (NF(3)) and optimal number of reference genes (NF(opt)). 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 NF(3) and NF(opt). Overall, the present study showed that HMBS, YWHAZ, SDHA, and TBP are suitable reference genes for qRT-PCR in porcine MSCs. Hindawi Publishing Corporation 2015 2015-04-20 /pmc/articles/PMC4417979/ /pubmed/25972899 http://dx.doi.org/10.1155/2015/235192 Text en Copyright © 2015 Won-Jae Lee et al. https://creativecommons.org/licenses/by/3.0/ 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.
spellingShingle Research Article
Lee, Won-Jae
Jeon, Ryoung-Hoon
Jang, Si-Jung
Park, Ji-Sung
Lee, Seung-Chan
Baregundi Subbarao, Raghavendra
Lee, Sung-Lim
Park, Bong-Wook
King, William Allan
Rho, Gyu-Jin
Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title_full Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title_fullStr Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title_full_unstemmed Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title_short Selection of Reference Genes for Quantitative Gene Expression in Porcine Mesenchymal Stem Cells Derived from Various Sources along with Differentiation into Multilineages
title_sort selection of reference genes for quantitative gene expression in porcine mesenchymal stem cells derived from various sources along with differentiation into multilineages
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4417979/
https://www.ncbi.nlm.nih.gov/pubmed/25972899
http://dx.doi.org/10.1155/2015/235192
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