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Identification and characterization of in vitro expanded hematopoietic stem cells

Che, James L C ; Bode, Daniel ; Kucinski, Iwo ; Cull, Alyssa H ; Bain, Fiona ; Becker, Hans J ; Jassinskaja, Maria LU ; Barile, Melania ; Boyd, Grace and Belmonte, Miriam , et al. (2022) In EMBO Reports 23(10).
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... (More)

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.

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Cells, Cultured, Endothelial Protein C Receptor/metabolism, Follistatin-Related Proteins/metabolism, Hematopoietic Stem Cells/metabolism, Humans, Mice, Transcription Factors/metabolism
in
EMBO Reports
volume
23
issue
10
article number
e55502
publisher
Springer Science and Business Media B.V.
external identifiers
  • pmid:35971894
  • scopus:85135960289
ISSN
1469-221X
DOI
10.15252/embr.202255502
language
English
LU publication?
no
additional info
© 2022 The Authors. Published under the terms of the CC BY 4.0 license.
id
c2f1f089-06c4-4f7e-ae93-c9e5f0d16dbb
date added to LUP
2025-01-16 16:04:00
date last changed
2025-07-18 19:12:03
@article{c2f1f089-06c4-4f7e-ae93-c9e5f0d16dbb,
  abstract     = {{<p>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 &gt; 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.</p>}},
  author       = {{Che, James L C and Bode, Daniel and Kucinski, Iwo and Cull, Alyssa H and Bain, Fiona and Becker, Hans J and Jassinskaja, Maria and Barile, Melania and Boyd, Grace and Belmonte, Miriam and Zeng, Andy G X and Igarashi, Kyomi J and Rubio-Lara, Juan and Shepherd, Mairi S and Clay, Anna and Dick, John E and Wilkinson, Adam C and Nakauchi, Hiromitsu and Yamazaki, Satoshi and Göttgens, Berthold and Kent, David G}},
  issn         = {{1469-221X}},
  keywords     = {{Animals; Cells, Cultured; Endothelial Protein C Receptor/metabolism; Follistatin-Related Proteins/metabolism; Hematopoietic Stem Cells/metabolism; Humans; Mice; Transcription Factors/metabolism}},
  language     = {{eng}},
  month        = {{10}},
  number       = {{10}},
  publisher    = {{Springer Science and Business Media B.V.}},
  series       = {{EMBO Reports}},
  title        = {{Identification and characterization of in vitro expanded hematopoietic stem cells}},
  url          = {{http://dx.doi.org/10.15252/embr.202255502}},
  doi          = {{10.15252/embr.202255502}},
  volume       = {{23}},
  year         = {{2022}},
}