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Related Experiment Video

Updated: May 21, 2026

Differentiation and Characterization of Neural Progenitors and Neurons from Mouse Embryonic Stem Cells
08:47

Differentiation and Characterization of Neural Progenitors and Neurons from Mouse Embryonic Stem Cells

Published on: May 15, 2020

Reference genes in the developing murine brain and in differentiating embryonic stem cells.

Nadine Kraemer1, Gerda Neubert, Lina Issa

  • 1Institute of Neuroanatomy and Cell Biology, Charité - Universitätsmedizin Berlin, Germany. nadine.kraemer@charite.de

Neurological Research
|June 28, 2012
PubMed
Summary

Selecting appropriate reference genes is crucial for accurate gene expression analysis. This study identifies RpII for mouse embryonic stem cell differentiation and Gapdh or Hprt for the developing murine neocortex.

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Area of Science:

  • Molecular Biology
  • Developmental Biology

Background:

  • Quantitative real-time PCR (qPCR) is essential for gene expression analysis in research.
  • Reference genes are used to normalize data, but their stability varies across experimental conditions and cell types.
  • Identifying stable reference genes is critical for accurate results, especially in developmental studies.

Purpose of the Study:

  • To identify suitable reference genes for gene expression analysis in the developing murine brain neocortex in vivo.
  • To find stable reference genes for mouse embryonic stem cells (mESC) during in vitro differentiation.

Main Methods:

  • Evaluated expression stability of five candidate genes (Actb, 18s, Gapdh, Hprt, RpII).
  • Analyzed gene expression in developing murine neocortex and differentiating mESC in vitro.
  • Utilized C(q) values, fold change, coefficient of variation (CV), and statistical measures to assess stability.

Main Results:

  • RpII demonstrated the highest expression stability in differentiating mESC.
  • Gapdh exhibited the greatest expression stability in the developing murine neocortex.
  • Candidate gene stability varied significantly between the two experimental systems.

Conclusions:

  • RpII is recommended as a reference gene for mESC differentiation studies.
  • Gapdh or Hprt are suggested as reference genes for murine neocortex neurodevelopment studies.
  • The choice of reference gene must be validated for specific experimental contexts in neurodevelopment research.