Abstract
SummaryHeterogeneity 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.
Highlights
Hematopoiesis is one of the best described models of adult stem cell biology due to the accessibility of tissue and the ability to isolate and functionally characterize multiple stages of a clearly defined hierarchy of differentiation (Bryder et al, 2006; Ema et al, 2014)
To gain deeper insight into the gene expression program of murine hematopoietic stem cells (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
Summary
Hematopoiesis is one of the best described models of adult stem cell biology due to the accessibility of tissue and the ability to isolate and functionally characterize multiple stages of a clearly defined hierarchy of differentiation (Bryder et al, 2006; Ema et al, 2014). Single-cell and serial transplantation studies have revealed significant heterogeneity in both the mature cell production and self-renewal durability of individual HSCs (Beerman et al, 2010; Dykstra et al, 2007; Goodell et al, 1996; Morita et al, 2010). This functional heterogeneity is thought to be controlled via cell intrinsic and extrinsic mechanisms (Copley and Eaves, 2013; Wilkinson and Gottgens, 2013) and is thought to play a role in disease evolution (Prick et al, 2014). While this has led to the identification of dozens of markers that enrich for HSC populations containing long-term HSCs (LT-HSCs), it is unclear which cells are HSCs and which are contaminating cells within any given HSC-enriched population
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
