Reliable reference genes for expression analysis of proliferating and adipogenically differentiating human adipose stromal cells

Cellular & Molecular Biology Letters
Claudia KrautgasserWerner Zwerschke

Abstract

The proliferation and adipogenic differentiation of adipose stromal cells (ASCs) are complex processes comprising major phenotypical alterations driven by up- and downregulation of hundreds of genes. Quantitative RT-PCR can be employed to measure relative changes in the expression of a gene of interest. This approach requires constitutively expressed reference genes for normalization to counteract inter-sample variations due to differences in RNA quality and quantity. Thus, a careful validation of quantitative RT-PCR reference genes is needed to accurately measure fluctuations in the expression of genes. Here, we evaluated candidate reference genes applicable for quantitative RT-PCR analysis of gene expression during proliferation and adipogenesis of human ASCs with the immunophenotype DLK1+/CD34+/CD90+/CD105+/CD45-/CD31-. We evaluated the applicability of 10 candidate reference genes (GAPDH, TBP, RPS18, EF1A, TFRC, GUSB, PSMD5, CCNA2, LMNA and MRPL19) using NormFinder, geNorm and BestKeeper software. The results indicate that EF1A and MRPL19 are the most reliable reference genes for quantitative RT-PCR analysis of proliferating ASCs. PSMD5 serves as the most reliable endogenous control in adipogenesis. CCNA2 and LMNA were amon...Continue Reading

References

Aug 3, 2005·BioTechniques·Marisa L Wong, Juan F Medrano
Oct 3, 2006·Cell Metabolism·Stephen R Farmer
Jun 4, 2008·Annual Review of Biochemistry·Peter Tontonoz, Bruce M Spiegelman
Sep 20, 2008·Science·Wei TangJonathan M Graff
Oct 7, 2008·Cell·Matthew S RodehefferJeffrey M Friedman
Nov 4, 2008·BMC Bioinformatics·Samuel ArvidssonBernd Mueller-Roeber
Mar 20, 2009·Cell Motility and the Cytoskeleton·Davina TondeleirAnja Lambrechts
Jan 25, 2011·Nature Reviews. Cancer·Rob A CairnsTak W Mak
Sep 9, 2011·Cold Spring Harbor Symposia on Quantitative Biology·R Siersbæk, S Mandrup
Sep 29, 2011·Nature Reviews. Molecular Cell Biology·Ana G Cristancho, Mitchell A Lazar
May 17, 2012·Theory in Biosciences = Theorie in Den Biowissenschaften·Jitesh Dundas, Maurice Ling
Jul 24, 2013·Nature Reviews. Molecular Cell Biology·Cosetta BertoliRobertus A M de Bruin
Sep 6, 2015·Stem Cell Research·Marit E ZwierzinaWerner Zwerschke
Jun 6, 2016·Trends in Endocrinology and Metabolism : TEM·Daniel C BerryJonathan M Graff
Nov 8, 2016·Biochemical and Biophysical Research Communications·Dylan LiLi Qiang
Oct 14, 2017·Aesthetic Surgery Journal·Ryan A LockhartSeveriano Dos-Anjos Vilaboa

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Methods Mentioned

BETA
FCS
Fluorescence-activated cell sorting
FACS
electrophoresis
Protein Assay
flow cytometry
PCR

Software Mentioned

geNorm
normfinder
BestKeeper
Flowing
GeNorm NormFinder
GraphPad Prism

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