RPL13A and EEF1A1 Are Suitable Reference Genes for qPCR during Adipocyte Differentiation of Vascular Stromal Cells from Patients with Different BMI and HOMA-IR.
Adriana-Mariel Gentile, Said Lhamyani, Leticia Coín-Aragüez, Wilfredo Oliva-Olivera, Hatem Zayed, Antonio Vega-Rioja, Javier Monteseirin, Silvana-Yanina Romero-Zerbo, Francisco-José Tinahones, Francisco-Javier Bermúdez-Silva, Rajaa El Bekay
Author Information
Adriana-Mariel Gentile: IBIMA, Universidad de Málaga, Campus Teatinos s/n, 29010, Málaga, España.
Said Lhamyani: IBIMA, Universidad de Málaga, Campus Teatinos s/n, 29010, Málaga, España.
Leticia Coín-Aragüez: CIBER Pathophysiology of obesity and nutrition CB06/03, Carlos III Health Institute, Malaga, 29010, Spain, Laboratory of Biomedical Research, Virgen de la Victoria Clinical University Hospital, Málaga, 29010, Spain.
Wilfredo Oliva-Olivera: CIBER Pathophysiology of obesity and nutrition CB06/03, Carlos III Health Institute, Malaga, 29010, Spain, Laboratory of Biomedical Research, Virgen de la Victoria Clinical University Hospital, Málaga, 29010, Spain.
Hatem Zayed: Biomedical Sciences Program, Health Sciences Department, College of Arts and Sciences, Qatar University, P.O. Box 2713, Doha, Qatar.
Antonio Vega-Rioja: Unidad de Gestión Clínica de Alergia Intercentros, Hospital Universitario Virgen Macarena, Sevilla, Spain.
Javier Monteseirin: Unidad de Gestión Clínica de Alergia Intercentros, Hospital Universitario Virgen Macarena, Sevilla, Spain.
Silvana-Yanina Romero-Zerbo: Unidad de Gestion Clínica Intercentros de Endocrinología y Nutrición, Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital Regional Universitario de Málaga/Universidad de Málaga, 29009, Málaga, Spain.
Francisco-José Tinahones: CIBER Pathophysiology of obesity and nutrition CB06/03, Carlos III Health Institute, Malaga, 29010, Spain, Laboratory of Biomedical Research, Virgen de la Victoria Clinical University Hospital, Málaga, 29010, Spain.
Francisco-Javier Bermúdez-Silva: Unidad de Gestion Clínica Intercentros de Endocrinología y Nutrición, Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital Regional Universitario de Málaga/Universidad de Málaga, 29009, Málaga, Spain.
Rajaa El Bekay: CIBER Pathophysiology of obesity and nutrition CB06/03, Carlos III Health Institute, Malaga, 29010, Spain, Laboratory of Biomedical Research, Virgen de la Victoria Clinical University Hospital, Málaga, 29010, Spain.
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.
