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Evaluation of suitable reference gene for real-time PCR in human umbilical cord mesenchymal stem cells with long-term in vitro expansion

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Abstract

In real-time quantitative PCR, the accuracy of normalized data is highly dependent on the stability of the reference genes. However, reference gene expression in a given cell type or experimental condition can vary considerably. The goal of this study was to establish a reliable set of reference genes for real-time PCR studies using human umbilical cord mesenchymal stem cells with long-term in vitro expansion. The stability of ten potential reference genes was examined in human umbilical cord mesenchymal stem cells. We found that Ywhaz and Rpl13a, not beta-actin or Gapdh, were the most stably expressed of the internal control genes in different passages of human umbilical cord mesenchymal stem cells. Ywhaz and Rpl13a could be used as reference genes for relative gene quantification and normalization purposes in real-time PCR studies of human umbilical cord mesenchymal stem cells.

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Acknowledgments

This study was supported by 863 projects from Ministry Science & Technology of China (2006AA02A110), National Natural Science Foundation of China (30600238).

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Authors and Affiliations

  1. National Engineering Research Center of Cell Products, AmCellGene Co. Ltd, TEDA, Tianjin, 300457, People’s Republic of China

    Youwei Wang, Shulin Yan, Aibin Mao, Bin Wang, He Ren & Zhongchao Han

  2. State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, National Engineering Research Center of Cell Products, 288 Nanjing Road, Tianjin, 300020, People’s Republic of China

    Zhibo Han, Ying Chi & Zhongchao Han

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  1. Youwei Wang
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  2. Zhibo Han
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  3. Shulin Yan
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  4. Aibin Mao
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  5. Bin Wang
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  6. He Ren
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  7. Ying Chi
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  8. Zhongchao Han
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Corresponding author

Correspondence to Zhongchao Han.

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Editor: J. Denry Sato

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Supplementary Figure 1

Expression stability and housekeeping gene ranking based on geNorm calculations. The M values of reference genes of UC-MSCs at passage 7 are calculated. (DOC 38 kb)

Supplementary Figure 2

Expression stability and housekeeping gene ranking based on geNorm calculations. The M values of reference genes of UC-MSCs at passage 15 are calculated. (DOC 54 kb)

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Wang, Y., Han, Z., Yan, S. et al. Evaluation of suitable reference gene for real-time PCR in human umbilical cord mesenchymal stem cells with long-term in vitro expansion. In Vitro Cell.Dev.Biol.-Animal 46, 595–599 (2010). https://doi.org/10.1007/s11626-010-9318-y

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  • DOI: https://doi.org/10.1007/s11626-010-9318-y

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