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Real-time or quantitative PCR (qPCR) is a useful technique that requires reliable reference genes for data normalization in gene expression analysis. Adipogenesis is among the biological processes suitable for this technique. The selection of adequate reference genes is essential for qPCR gene expression analysis of human Vascular Stromal Cells (hVSCs) during their differentiation into adipocytes. To the best of our knowledge, there are no studies validating reference genes for the analyses of visceral and subcutaneous adipose tissue hVSCs from subjects with different Body Mass Index (BMI) and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) index. The present study was undertaken to analyze this question. We first analyzed the stability of expression of five potential reference genes:  CYC ,  GAPDH ,  RPL13A ,  EEF1A1 , and  18S  ribosomal RNA, during  in vitro  adipogenic differentiation, in samples from these types of patients. The expression of  RPL13A  and  EEF1A1  was not affected by differentiation, thus being these genes the most stable candidates, while  CYC ,  GAPDH , and  18S  were not suitable for this sort of analysis. This work highlights that  RPL13A  and  EEF1A1  are good candidates as reference genes for qPCR analysis of hVSCs differentiation into adipocytes from subjects with different BMI and HOMA-IR.

plos one

RPL13A and EEF1A1 Are Suitable Reference Genes for qPCR during Adipocyte Differentiation of Vascular Stromal Cells from Patients with Different BMI and HOMA-IR