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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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Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations.

Nicola K Wilson1, David G Kent1, Florian Buettner2

  • 1Department of Haematology, Wellcome Trust and MRC Cambridge Stem Cell Institute and Cambridge Institute for Medical Research, Cambridge University, Cambridge CB2 0XY, UK.

Cell Stem Cell
|May 26, 2015
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Summary

Researchers identified key molecules driving durable self-renewal in hematopoietic stem cells (HSCs). This single-cell approach links molecular data with functional stem cell activity for better understanding of HSC function.

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Area of Science:

  • Stem cell biology
  • Hematopoiesis
  • Molecular biology

Background:

  • Adult hematopoietic stem cells (HSCs) exhibit heterogeneity in self-renewal durability, complicating studies of their function.
  • Gene expression studies are hindered by mixed HSC subtypes and non-HSC contaminants in bulk populations.

Purpose of the Study:

  • To gain deeper insight into the gene expression program of murine HSCs.
  • To identify key molecules associated with long-term durable self-renewal.
  • To develop an unbiased sorting strategy for HSCs.

Main Methods:

  • Combined single-cell functional assays with flow cytometric index sorting.
  • Utilized single-cell gene expression assays (RNA-seq).
  • Integrated bioinformatic analysis of datasets for unbiased cell sorting and molecular identification.

Main Results:

  • Developed an unbiased sorting strategy to separate HSCs from non-HSCs.
  • Identified key molecules linked to long-term durable HSC self-renewal.
  • Created a single-cell molecular dataset correlated with functional stem cell activity.

Conclusions:

  • The developed single-cell approach effectively links molecular profiles with functional stem cell activity.
  • This methodology can be applied to other stem cell systems for identifying functional molecules.