Rapid myeloerythroid repopulation after intrafemoral transplantation of NOD-SCID mice reveals a new class of human stem cells

Abstract

A major problem hampering effective stem cell–based therapies is the absence of a clear understanding of the human hematopoietic stem cell (HSC) pool composition. The severe combined immunodeficiency (SCID) repopulating cell (SRC) xenotransplant assay system provides a powerful tool for characterizing the frequency, cell surface markers, cell cycle status, homing and response to cytokine stimulation of human HSCs1,2,3. Clonal tracking of retrovirally transduced SRCs and transplantation of specific subpopulations revealed SRC classes with distinct repopulation potentials4,5,6,7. However, all HSC repopulation assays are based on intravenous injection, a complex process that requires circulation through blood, recognition and extravasation through bone marrow vasculature, and migration to a supportive microenvironment8,9,10,11. Thus, some classes of HSCs may remain undetected. By direct intrafemoral injection, we identified rapid SRCs (R-SRCs) within the LinCD34+CD38loCD36 subpopulation. R-SRCs rapidly generate high levels of human myeloid and erythroid cells within the injected femur, migrate to the blood and colonize individual bones of non-obese diabetic (NOD)-SCID mice within 2 weeks after transplantation. Lentivector-mediated clonal analysis of individual R-SRCs revealed heterogeneity in their proliferative and migratory properties. The identification of a new HSC class and an effective intrafemoral assay provide the tools required to develop more effective stem cell–based therapies that rely on rapid reconstitution.

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Figure 1: Kinetic analysis of human cell engraftment after intrafemoral and intravenous delivery of LinCD34+ cord blood cells.
Figure 2: Multilineage engraftment of human cells after intrafemoral transplantation into NOD-SCID mice.
Figure 3: R-SRCs belong to the LinCD34+CD38loCD36 cell fraction.
Figure 4: Heterogeneity of the proliferative and migration potential of gene-marked cells 3 weeks after intrafemoral injection.

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Acknowledgements

We thank P Scheufler, P. Savage and the entire Obstetrics unit (Trillium Hospital) for providing cord blood samples; J. McKenzie for assistance with Southern analysis; S. Zhao (Hospital for Sick Children) for sorting; and T. Lapidot (Wiezmann Institute) and members of the Dick lab for critical comments on the manuscript. This work was supported by grants from the Association pour la Recherche contre le Cancer (F.M.), the Stem Cell Network of National Centres of Excellence (F.M. and J.E.D.) and the National Cancer Institute of Canada, with funds from the Canadian Cancer Society, the Canadian Genetic Diseases Network of the National Centres of Excellence, the Canadian Institutes for Health Research and a Canada Research Chair (J.E.D.).

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Correspondence to John E Dick.

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Mazurier, F., Doedens, M., Gan, O. et al. Rapid myeloerythroid repopulation after intrafemoral transplantation of NOD-SCID mice reveals a new class of human stem cells. Nat Med 9, 959–963 (2003). https://doi.org/10.1038/nm886

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