Evaluation of suitable reference gene for real-time PCR in human umbilical cord mesenchymal stem cells with long-term in vitro expansion.
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.
Wang
Youwei
Y
National Engineering Research Center of Cell Products, AmCellGene Co. Ltd, TEDA, Tianjin 300457, People's Republic of China.
Han
Zhibo
Z
Yan
Shulin
S
Mao
Aibin
A
Wang
Bin
B
Ren
He
H
Chi
Ying
Y
Han
Zhongchao
Z
D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D048430:Cell Shape; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D005796:Genes / Q000235:genetics*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012984:Software; D013997:Time Factors; D014470:Umbilical Cord / Q000166:cytology*