PMID- 18976469
OWN - NLM
STAT- MEDLINE
DCOM- 20081211
LR  - 20220408
IS  - 1471-2199 (Electronic)
IS  - 1471-2199 (Linking)
VI  - 9
DP  - 2008 Oct 31
TI  - Instability of standard PCR reference genes in adipose-derived stem cells during 
      propagation, differentiation and hypoxic exposure.
PG  - 98
LID - 10.1186/1471-2199-9-98 [doi]
AB  - BACKGROUND: For the accurate determination of gene expression changes during 
      growth and differentiation studies on adipose-derived stem cells (ASCs), 
      quantitative real-time RT-PCR has become a method of choice. The technology is 
      very sensitive, however, without a proper selection of reference genes, to which 
      the genes of interest are normalized, erroneous results may be obtained. RESULTS: 
      In this study, we have compared the gene expression levels of a panel of twelve 
      widely used reference genes during hypoxic culture, osteogenic and chondrogenic 
      differentiation, and passaging of primary human ASCs. We found that several of 
      the commonly used reference genes including 18S rRNA, glyceraldehyde-3-phosphate 
      dehydrogenase (GAPDH) and beta-actin were unsuitable for normalization in the 
      conditions we tested, whereas tyrosine 3/tryptophan 5-monooxygenase activation 
      protein (YMHAZ), TATAA-box binding protein (TBP), beta-glucuronidase (GUSB) were 
      the most stable across all conditions. CONCLUSION: When determining gene 
      expression levels in adipose-derived stem cells, we recommend normalizing 
      transcription levels to the geometric mean of YMHAZ, TBP and GUSB.
FAU - Fink, Trine
AU  - Fink T
AD  - Laboratory for Stem Cell Research, Aalborg University, Denmark. trinef@hst.aau.dk
FAU - Lund, Pia
AU  - Lund P
FAU - Pilgaard, Linda
AU  - Pilgaard L
FAU - Rasmussen, Jeppe Grondahl
AU  - Rasmussen JG
FAU - Duroux, Meg
AU  - Duroux M
FAU - Zachar, Vladimir
AU  - Zachar V
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20081031
PL  - England
TA  - BMC Mol Biol
JT  - BMC molecular biology
JID - 100966983
RN  - 0 (Actins)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - 63231-63-0 (RNA)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
RN  - EC 3.2.1.31 (Glucuronidase)
SB  - IM
MH  - Actins/genetics
MH  - Adipose Tissue/*cytology
MH  - Adult
MH  - *Cell Differentiation
MH  - Chondrogenesis
MH  - Female
MH  - *Gene Expression Profiling
MH  - Glucuronidase/genetics
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
MH  - Humans
MH  - Hypoxia/*genetics
MH  - Mesenchymal Stem Cells/chemistry/*cytology
MH  - Middle Aged
MH  - Osteogenesis
MH  - Polymerase Chain Reaction/*standards
MH  - RNA/analysis
MH  - RNA, Ribosomal, 18S/genetics
MH  - Reference Standards
PMC - PMC2585587
EDAT- 2008/11/04 09:00
MHDA- 2008/12/17 09:00
PMCR- 2008/10/31
CRDT- 2008/11/04 09:00
PHST- 2008/05/24 00:00 [received]
PHST- 2008/10/31 00:00 [accepted]
PHST- 2008/11/04 09:00 [pubmed]
PHST- 2008/12/17 09:00 [medline]
PHST- 2008/11/04 09:00 [entrez]
PHST- 2008/10/31 00:00 [pmc-release]
AID - 1471-2199-9-98 [pii]
AID - 10.1186/1471-2199-9-98 [doi]
PST - epublish
SO  - BMC Mol Biol. 2008 Oct 31;9:98. doi: 10.1186/1471-2199-9-98.