Abstract:
Adipose-derived stromal cells (ASCs) are becoming increasingly attractive as cellular therapy products. Their
differentiation potential, the secretion of growth and differentiation factors, and the ability to cryopreserve the cells
over extended periods are important features. Changes in experimental conditions result in changes in gene expression, and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) has become an important tool
for measuring these changes. There is, however, the potential to introduce technical bias in the process, which can be
diminished through the selection of stable reference genes (RGs). Using geNorm software, in this in vitro study we
explore the effects that adipogenic differentiation for a 21 day induction period, cryopreservation (freshly isolated
ASCs or previously cryopreserved/frozen ASCs), and culture medium supplementation (fetal bovine serum vs.
pooled human platelet lysate) have on the stability of 11 RGs. We found that RG stability is markedly affected by the
different experimental conditions. Of the RGs assessed, YWHAZ, HPRT, TBP, andACTB were stably expressed genes
under all experimental conditions. We recommend that a panel of stable RGs should be selected before studying gene
expression during adipogenesis, and that this is based on the experimental condition(s) being investigated.
