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Background  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 − .    Methods  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.    Results  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 among the least consistent genes.    Conclusions  Applying these findings for future gene expression analyses will help elucidate ASC biology.    Electronic supplementary material  The online version of this article (10.1186/s11658-019-0140-6) contains supplementary material, which is available to authorized users.

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