Che, James L C, Bode, Daniel, Kucinski, Iwo et al. (18 more authors) (2022) Identification and characterization of in vitro expanded hematopoietic stem cells. EMBO Reports. e55502. e55502. ISSN 1469-221X
Abstract
Hematopoietic stem cells (HSCs) cultured outside the body are the fundamental component of a wide range of cellular and gene therapies. Recent efforts have achieved > 200-fold expansion of functional HSCs, but their molecular characterization has not been possible since the majority of cells are non-HSCs and single cell-initiated cultures have substantial clone-to-clone variability. Using the Fgd5 reporter mouse in combination with the EPCR surface marker, we report exclusive identification of HSCs from non-HSCs in expansion cultures. By directly linking single-clone functional transplantation data with single-clone gene expression profiling, we show that the molecular profile of expanded HSCs is similar to proliferating fetal HSCs and reveals a gene expression signature, including Esam, Prdm16, Fstl1, and Palld, that can identify functional HSCs from multiple cellular states. This "repopulation signature" (RepopSig) also enriches for HSCs in human datasets. Together, these findings demonstrate the power of integrating functional and molecular datasets to better derive meaningful gene signatures and opens the opportunity for a wide range of functional screening and molecular experiments previously not possible due to limited HSC numbers.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2022The Authors |
Keywords: | Animals,Cells, Cultured,Endothelial Protein C Receptor/metabolism,Follistatin-Related Proteins/metabolism,Hematopoietic Stem Cells/metabolism,Humans,Mice,Transcription Factors/metabolism |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) |
Depositing User: | Pure (York) |
Date Deposited: | 01 Aug 2022 14:30 |
Last Modified: | 27 Dec 2024 00:21 |
Published Version: | https://doi.org/10.15252/embr.202255502 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.15252/embr.202255502 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:189611 |
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Description: Identification and characterization of in vitro expanded hematopoietic stem cells
Licence: CC-BY 2.5