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Updated: May 16, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
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Sorting live stem cells based on Sox2 mRNA expression.

Hans M Larsson1, Seung Tae Lee, Marta Roccio

  • 1Laboratory for Regenerative Medicine and Pharmacobiology, Institute for Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

Plos One
|December 5, 2012
PubMed
Summary
This summary is machine-generated.

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Molecular beacons (MBs) enable sorting stem cells using mRNA markers, not just surface proteins. This technique successfully purified undifferentiated stem cells and enhanced neurosphere generation from sorted neural stem cells.

Area of Science:

  • Biotechnology
  • Stem Cell Biology
  • Molecular Biology

Background:

  • Traditional cell sorting relies on cell-surface protein markers.
  • Molecular beacons (MBs) offer a novel approach to sort cells based on intracellular mRNA expression.
  • This method holds potential for isolating rare cell populations.

Purpose of the Study:

  • To demonstrate the utility of MBs for purifying stem cells based on mRNA expression.
  • To develop and validate MBs targeting the Sox2 mRNA, a key stem cell marker.
  • To assess the efficiency of MB-based sorting for stem cell isolation and differentiation.

Main Methods:

  • Design and synthesis of molecular beacons targeting Sox2 mRNA.
  • In vitro and in vivo validation of MB fluorescence in murine embryonic stem cells (mES) and neural stem cells (NSCs).

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Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
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  • Cell sorting of Sox2-MB(+)SSEA1(+) cells from mixed mES cell populations and Sox2-MB(+) cells from primary mouse brain isolates.

    • Main Results:

    • A designed Sox2-targeting MB showed increased fluorescence in mES and NSCs compared to a nonspecific MB.
    • Live, undifferentiated stem cells were effectively isolated from mixed mES cell populations using Sox2 MBs.
    • Sox2-MB(+) cells sorted from primary mouse brains exhibited higher neurosphere generation efficiency than Sox2-MB(-) cells.

    Conclusions:

    • Molecular beacons provide a powerful tool for mRNA-specific stem cell sorting.
    • This technique enables the isolation of specific stem cell populations based on gene expression.
    • MB-based sorting enhances the purification and functional assessment of stem cells.