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

Wilson NK, Kent DG, Buettner F, Shehata M, Macaulay IC, Calero-Nieto FJ, Castillo MS, Oedekoven CA, Diamanti E, Schulte R, Ponting CP, Voet T, Caldas C, Stingl J, Green AR, Theis FJ, Goettgens B (2015)

Publication Type: Journal article

Publication year: 2015

Journal

Cell Stem Cell Elsevier (Cell Press)

Book Volume: 16

Pages Range: 712-724

Journal Issue: 6

DOI: 10.1016/j.stem.2015.04.004

Abstract

Heterogeneity within the self-renewal durability of adult hematopoietic stem cells (HSCs) challenges our understanding of the molecular framework underlying HSC function. Gene expression studies have been hampered by the presence of multiple HSC subtypes and contaminating non-HSCs in bulk HSC populations. To gain deeper insight into the gene expression program of murine HSCs, we combined single-cell functional assays with flow cytometric index sorting and single-cell gene expression assays. Through bioinformatic integration of these datasets, we designed an unbiased sorting strategy that separates non-HSCs away from HSCs, and single-cell transplantation experiments using the enriched population were combined with RNA-seq data to identify key molecules that associate with long-term durable self-renewal, producing a single-cell molecular dataset that is linked to functional stem cell activity. Finally, we demonstrated the broader applicability of this approach for linking key molecules with defined cellular functions in another stem cell system.

Involved external institutions

University of Cambridge GB United Kingdom (GB) Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (HMGU) / Helmholtz Munich DE Germany (DE) Wellcome Trust Sanger Institute - Genome Research Limited GB United Kingdom (GB) University of Oxford GB United Kingdom (GB)

How to cite

APA:

Wilson, N.K., Kent, D.G., Buettner, F., Shehata, M., Macaulay, I.C., Calero-Nieto, F.J.,... Goettgens, B. (2015). Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations. Cell Stem Cell, 16(6), 712-724. https://doi.org/10.1016/j.stem.2015.04.004

MLA:

Wilson, Nicola K., et al. "Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations." Cell Stem Cell 16.6 (2015): 712-724.

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