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Identification and characterization of in vitro expanded hematopoietic stem cells
Author(s) -
Che James L C,
Bode Daniel,
Kucinski Iwo,
Cull Alyssa H,
Bain Fiona,
Becker Hans J,
Jassinskaja Maria,
Barile Melania,
Boyd Grace,
Belmonte Miriam,
Zeng Andy G X,
Igarashi Kyomi J,
RubioLara Juan,
Shepherd Mairi S,
Clay Anna,
Dick John E,
Wilkinson Adam C,
Nakauchi Hiromitsu,
Yamazaki Satoshi,
Göttgens Berthold,
Kent David G
Publication year - 2022
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.15252/embr.202255502
Subject(s) - biology , clone (java method) , haematopoiesis , stem cell , gene expression profiling , microbiology and biotechnology , gene , computational biology , transplantation , genetics , gene expression , medicine , surgery
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.