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Reference Gene Validation via RT--qPCR for Human iPSC-Derived Neural Stem Cells and Neural Progenitors

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cris.lastimport.scopus2024-02-12T19:47:21Z
dc.abstract.enCorrect 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 $Δ$Ct 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.
dc.affiliationUniwersytet Warszawski
dc.contributor.authorLenart, Jacek
dc.contributor.authorLipka, Gabriela
dc.contributor.authorAugustyniak, Justyna
dc.contributor.authorBużańska, Leonora
dc.contributor.authorStępień, Piotr
dc.date.accessioned2024-01-25T19:08:47Z
dc.date.available2024-01-25T19:08:47Z
dc.date.copyright2019-03-29
dc.date.issued2019
dc.description.accesstimeBEFORE_PUBLICATION
dc.description.financeNie dotyczy
dc.description.versionFINAL_PUBLISHED
dc.description.volume56
dc.identifier.doi10.1007/S12035-019-1538-X
dc.identifier.issn0893-7648
dc.identifier.urihttps://repozytorium.uw.edu.pl//handle/item/118132
dc.identifier.weblinkhttps://link.springer.com/article/10.1007%2Fs12035-019-1538-x
dc.languageeng
dc.pbn.affiliationbiological sciences
dc.relation.ispartofMolecular Neurobiology
dc.relation.pages6820–6832
dc.rightsCC-BY
dc.sciencecloudnosend
dc.subject.enhuman induced Pluripotent Stem Cell (hiPSC)
dc.subject.enNeural Stem Cells
dc.subject.enNeural Progenitor
dc.subject.enquantitative real-time Polymerase Chain Reaction (RT-qPCR)
dc.subject.enGene expression reference panel
dc.subject.enRelative gene expression
dc.titleReference Gene Validation via RT--qPCR for Human iPSC-Derived Neural Stem Cells and Neural Progenitors
dc.typeJournalArticle
dspace.entity.typePublication