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

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 popula...

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Autores principales: Wilson, Nicola K., Kent, David G., Buettner, Florian, Shehata, Mona, Macaulay, Iain C., Calero-Nieto, Fernando J., Sánchez Castillo, Manuel, Oedekoven, Caroline A., Diamanti, Evangelia, Schulte, Reiner, Ponting, Chris P., Voet, Thierry, Caldas, Carlos, Stingl, John, Green, Anthony R., Theis, Fabian J., Göttgens, Berthold
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460190/
https://www.ncbi.nlm.nih.gov/pubmed/26004780
http://dx.doi.org/10.1016/j.stem.2015.04.004
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author Wilson, Nicola K.
Kent, David G.
Buettner, Florian
Shehata, Mona
Macaulay, Iain C.
Calero-Nieto, Fernando J.
Sánchez Castillo, Manuel
Oedekoven, Caroline A.
Diamanti, Evangelia
Schulte, Reiner
Ponting, Chris P.
Voet, Thierry
Caldas, Carlos
Stingl, John
Green, Anthony R.
Theis, Fabian J.
Göttgens, Berthold
author_facet Wilson, Nicola K.
Kent, David G.
Buettner, Florian
Shehata, Mona
Macaulay, Iain C.
Calero-Nieto, Fernando J.
Sánchez Castillo, Manuel
Oedekoven, Caroline A.
Diamanti, Evangelia
Schulte, Reiner
Ponting, Chris P.
Voet, Thierry
Caldas, Carlos
Stingl, John
Green, Anthony R.
Theis, Fabian J.
Göttgens, Berthold
author_sort Wilson, Nicola K.
collection PubMed
description 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.
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spelling pubmed-44601902015-06-16 Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations Wilson, Nicola K. Kent, David G. Buettner, Florian Shehata, Mona Macaulay, Iain C. Calero-Nieto, Fernando J. Sánchez Castillo, Manuel Oedekoven, Caroline A. Diamanti, Evangelia Schulte, Reiner Ponting, Chris P. Voet, Thierry Caldas, Carlos Stingl, John Green, Anthony R. Theis, Fabian J. Göttgens, Berthold Cell Stem Cell Resource 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. Cell Press 2015-06-04 /pmc/articles/PMC4460190/ /pubmed/26004780 http://dx.doi.org/10.1016/j.stem.2015.04.004 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Resource
Wilson, Nicola K.
Kent, David G.
Buettner, Florian
Shehata, Mona
Macaulay, Iain C.
Calero-Nieto, Fernando J.
Sánchez Castillo, Manuel
Oedekoven, Caroline A.
Diamanti, Evangelia
Schulte, Reiner
Ponting, Chris P.
Voet, Thierry
Caldas, Carlos
Stingl, John
Green, Anthony R.
Theis, Fabian J.
Göttgens, Berthold
Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title_full Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title_fullStr Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title_full_unstemmed Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title_short Combined Single-Cell Functional and Gene Expression Analysis Resolves Heterogeneity within Stem Cell Populations
title_sort combined single-cell functional and gene expression analysis resolves heterogeneity within stem cell populations
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460190/
https://www.ncbi.nlm.nih.gov/pubmed/26004780
http://dx.doi.org/10.1016/j.stem.2015.04.004
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