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Updated: Jan 27, 2026

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells
Published on: May 22, 2014
Reference Gene Validation via RT-qPCR for Human iPSC-Derived Neural Stem Cells and Neural Progenitors
Justyna Augustyniak1, Jacek Lenart2, Gabriela Lipka1
1Department of Stem Cell Bioengineering, Mossakowski Medical Research Centre Polish Academy of Sciences, 5 Pawinskiego Str., 02-106, Warsaw, Poland.
Abstract:
Correct selection of the reference gene(s) is the most important step in gene expression analysis. The aims of this study were to identify and evaluate the panel of possible reference genes in neural stem cells (NSC), early neural progenitors (eNP) and neural progenitors (NP) obtained from human-induced pluripotent stem cells (hiPSC). The stability of expression of genes commonly used as the reference in cells during neural differentiation is variable and does not meet the criteria for reference genes. In the present work, we evaluated the stability of expression of 16 candidate reference genes using the four most popular algorithms: the ΔCt method, BestKeeper, geNorm and NormFinder. All data were analysed using the online tool RefFinder to obtain a comprehensive ranking. Our results indicate that NormFinder is the best tool for reference gene selection in early stages of hiPSC neural differentiation. None of the 16 tested genes is suitable as reference gene for all three stages of development. We recommend using different genes (panel of genes) to normalise RT-qPCR data for each of the neural differentiation stages.
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