Methodological considerations on how to identify human hematopoietic stem cells

Hinchly, Taylor, Bonnet, Dominique and Anjos Afonso, Fernando ORCID: https://orcid.org/0000-0003-4392-488X 2025. Methodological considerations on how to identify human hematopoietic stem cells. Experimental Hematology: Journal for Hematology, Stem Cell Biology and Transplantation 144 , 104729. 10.1016/j.exphem.2025.104729

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Abstract

Recently, human CD34+ hematopoietic stem cells (HSCs) have been purified to a frequency of approximately 1 in 3 cells, a population denoted as CD34+CD38-CD45RA-CD90±EPCR+ HSCs. This work aimed to evaluate the methodology for CD34+ HSC isolation, exploring differences in antibody clones, conjugates, source of cells and additional cell surface antigens (integrin-α6, CLEC9A and GPRC5C) to enhance the purity of these EPCR+ HSCs. We are emphasizing here the importance of experimental planning and antibody panel selection concerning the isolation of these human HSCs from multiple sources and providing important notes on the pitfalls of the reagents used for such purposes. Our results should enable a better reproducibility of results between labs, as well as further pursue work towards improving the enrichment of human HSCs.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Schools > Biosciences
Publisher:	Elsevier
ISSN:	0301-472X
Funders:	The Jane Hodge Foundation, UK Biotechnology and Biological Sciences Research Council, Francis Crick Institute, Cancer Research UK, UK Medical Research Council, Wellcome Trust
Date of First Compliant Deposit:	31 January 2025
Date of Acceptance:	23 January 2025
Last Modified:	13 Mar 2025 13:55
URI:	https://orca.cardiff.ac.uk/id/eprint/175808

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