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A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations
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Single-cell mass cytometry analysis reveals stem cell heterogeneity.

Thulaj Meharwade1, Loïck Joumier1, Maxime Parisotto2

  • 1Montreal Clinical Research Institute (IRCM), 110 Avenue Des Pins Ouest, Montreal, QC H2W 1R7, Canada; Department of Biochemistry and Molecular Medicine, University of Montreal, C.P. 6128, Succursale Centre-ville, Montreal, QC H3C 3J7, Canada.

Methods (San Diego, Calif.)
|October 13, 2022
PubMed
Summary

Mass cytometry enables simultaneous single-cell protein analysis, revealing significant variations in stem cell regulators and cross-activated signaling pathways. This technology identifies distinct embryonic stem cell states and their variations under different culture conditions.

Keywords:
Cellular heterogeneityEmbryonic stem cellsMass cytometryPluripotent stem cellsSingle-cell protein analysiscytometry by time of flight (CyTOF)

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Area of Science:

  • Proteomics
  • Cell Biology
  • Developmental Biology

Background:

  • Cellular heterogeneity is crucial for development and disease.
  • Single-cell transcriptomics is advanced, but single-cell proteomics lags.
  • Understanding protein dynamics at the single-cell level is essential.

Purpose of the Study:

  • To develop and apply mass cytometry for simultaneous single-cell protein profiling.
  • To investigate protein level variations of key regulators in mouse embryonic stem cells.
  • To identify distinct cell states and signaling pathway activation in embryonic stem cells.

Main Methods:

  • Simultaneous multi-protein profiling using mass cytometry (cytometry by time of flight).
  • Development of mass cytometry reagents for transcription factors, signaling proteins, and chromatin modifiers.
  • Analysis of protein expression and cell states in mouse embryonic stem cells under various culture conditions.

Main Results:

  • Significant variation in protein levels of stem cell regulators was observed.
  • Extensive cross-activation of cell signaling pathways across different culture conditions.
  • Identification of distinct multiple cell states within embryonic stem cell populations.
  • Quantification of cell state variations influenced by culture conditions.

Conclusions:

  • Mass cytometry is a powerful tool for single-cell proteomic analysis.
  • Protein level heterogeneity is a key feature of embryonic stem cells.
  • This approach provides new insights into stem cell regulation and plasticity.
  • Future perspectives for single-cell protein analysis in developmental and disease contexts.