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Hematopoiesis
CD27, CD201, FLT3, CD48, and CD150 cell surface staining identifies long-term mouse hematopoietic stem cells in immunodeficient non-obese diabetic severe combined immune deficient-derived strains
Ferrata Storti Foundation
Haematologica 2020 Volume 105(1):71-82
Bianca Nowlan,1,2,3,4,5 Elizabeth D. Williams,2,4,5 Michael R. Doran,1,2,3,4,5,6,* and Jean-Pierre Levesque3,5,*
1Stem Cell Therapies Laboratory, School of Biomedical Science, Faculty of Health, Queensland University of Technology (QUT), Brisbane; 2School of Biomedical Science, Faculty of Health, Institute of Health and Biomedical Innovation, QUT, Kelvin Grove, Queensland; 3Mater Research Institute – The University of Queensland, Woolloongabba; 4Australian Prostate Cancer Research Centre - Queensland, Brisbane, Queensland; 5Translational Research Institute, Woolloongabba, Queensland and 6Australian National Centre for the Public Awareness of Science, Australian National University, Canberra, Australian Capital Territory, Australia
* MRD and JPL contributed equally to this work.
ABSTRACT
Staining for CD27 and CD201 (endothelial protein C receptor) has been recently suggested as an alternative to stem cell antigen–1 (Sca1) to identify hematopoietic stem cells in inbred mouse strains with low or nil expression of SCA1. However, whether staining for CD27 and CD201 is compatible with low fms-like tyrosine kinase 3 (FLT3) expression and the “SLAM” code defined by CD48 and CD150 to identify mouse long-term reconstituting hematopoietic stem cells has not been established. We com- pared the C57BL/6 strain, which expresses a high level of SCA1 on hematopoietic stem cells to non-obese diabetic severe combined immune deficient NOD.CB17-prkdcscid/Sz (NOD-scid) mice and NOD.CB17-prkdcscid il2rgtm1Wj1/Sz (NSG) mice which both express low to negative levels of SCA1 on hematopoietic stem cells. We demonstrate that hematopoietic stem cells are enriched within the linage-negative C-KIT+ CD27+ CD201+ FLT3- CD48- CD150+ population in serial dilution long-term competitive transplantation assays. We also make the novel observation that CD48 expression is up- regulated in Lin- KIT+ progenitors from NOD-scid and NSG strains, which otherwise have very few cells expressing the CD48 ligand CD244. Finally, we report that unlike hematopoietic stem cells, SCA1 expression is similar on bone marrow endothelial and mesenchymal progenitor cells in C57BL/6, NOD-scid and NSG mice. In conclusion, we propose that the combination of Lineage, KIT, CD27, CD201, FLT3, CD48, and CD150 antigens can be used to identify long-term reconstituting hematopoietic stem cells from mouse strains expressing low levels of SCA1 on hematopoietic cells.
Introduction
Blood myeloid and erythroid lineages are short-lived and require continuous replacement from hematopoietic stem cells (HSC) in the bone marrow (BM).1-6 HSC are defined by their capacity to clonally reconstitute the hematopoietic system in lethally irradiated mice upon transplantation. Using cell surface markers, mouse HSC are comprised within the LSK population of cells, i.e., cells negative for B, T, myeloid and erythroid lineages (Lin–), positive for c-KIT/CD117 and positive for stem cell antigen-1 (SCA1 or LY6A/E). Multipotent long-term reconstituting HSC (LT-HSC) are LSK cells that are negative for fms-like tyrosine kinase 3 (FLT3)/CD135 and CD48 and positive for signaling lymphocytic activation mole- cule (SLAMF1/CD150).4,5 When transplanted, these HSC can clonally and serially reconstitute hematopoiesis in lethally irradiated mice.5
Correspondence:
JEAN-PIERRE LEVESQUE
jp.levesque@mater.uq.edu.au
MICHAEL DORAN
mike@mikedoranlab.com
Received: November 22, 2018. Accepted: May 2, 2019. Pre-published: May 9, 2019.
doi:10.3324/haematol.2018.212910
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/105/1/71
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