Reliable Reference Genes for Expression Analysis of Proliferating and Adipogenically Differentiating Human Adipose Stromal Cells

Overview

Journal Cell Mol Biol Lett

Publisher Biomed Central

Specialties Biochemistry
Cell Biology
Molecular Biology

Date 2019 Mar 1

PMID 30815013

Citations 10

Authors

Claudia Krautgasser

Markus Mandl

Florian M Hatzmann

Petra Waldegger

Monika Mattesich

Werner Zwerschke

Affiliations

Soon will be listed here.

Abstract

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 (, , , , , , , , and ) using NormFinder, geNorm and BestKeeper software.

Results: The results indicate that and are the most reliable reference genes for quantitative RT-PCR analysis of proliferating ASCs. serves as the most reliable endogenous control in adipogenesis. and were among the least consistent genes.

Conclusions: Applying these findings for future gene expression analyses will help elucidate ASC biology.

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