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Distinct classes of human stem cells that differ in proliferative and self-renewal potential

Nature Immunology volume 2, pages 75–82 (2001)Cite this article

Abstract

The composition of the human hematopoietic stem cell compartment is poorly understood due to the absence of experimental tools with which to characterize the developmental program of individual stem cells. We report here that human stem cells differ markedly in their repopulation capacity and self-renewal potential, as determined using nonobese diabetic–severe combined immunodeficiency (NOD-SCID) mice transplanted with retrovirally transduced cord blood stem cells, called SCID-repopulating cells (SRCs). Clonal stem cell analysis based on the identification of unique retroviral integration sites within serial bone marrow aspirates showed that repopulation was generally oligoclonal with extensive variability in the lifespan and proliferative capacity of individual SRCs. Most clones contributed to human cell engraftment for several weeks after transplantation and then disappeared but others appeared later and persisted. Further evidence for stem cell heterogeneity was found in the secondary transplantation capacity of SRCs. These data point to the existence of different classes of human stem cells with variable self-renewal potential and short- or long-term repopulating capacity.

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Figure 1: Efficiency of gene transfer into Lin− CB cells and the kinetics of their repopulation of NOD-SCID mice.
Figure 2: Multilineage repopulation capacity of transduced SRCs.
Figure 3: Lymphohematopoietic distribution of total human and EGFP+ human cells in NOD-SCID mice.
Figure 4: Kinetic variation in the contribution of gene-marked cells to the human graft.
Figure 5: Southern blot analysis for provirus integration sites within the BM, spleen and thymus of mice engrafted with transduced human cells.
Figure 6: Distribution of the number of transduced clones per graft and the calculated cellularity of transduced clones.
Figure 7: Self-renewal capacity of transduced SRCs.

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Acknowledgements

We thank C. Botsford for the CB samples, G. Knowles for flow sorting of MGIN cells, R. G. Hawley and T. Hawley for the PG13-MGIN producer cell line, R. McInnes for the primers and template required for human chx10 sequence amplification and J. Ellis and R. McInnes for critical review of the manuscript. Supported by grants from the National Cancer Institute of Canada with funds from the Canadian Cancer Society, the Canadian Genetic Diseases Network of the National Centers of Excellence (to J. E. D.), the Medical Research Council (to J. E. D.), a Medical Research Council Scientist award (to J. E. D.) and the Comisión Interministerial de Ciencia y Tecnología (to G. G., grant number SAF9808C04-1).

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  1. Guillermo Guenechea, Olga I. Gan and Craig Dorrell: These authors contributed equally to this work.

Authors and Affiliations

  1. Programs in Cancer/Blood Research and Gene Therapy Hospital for Sick Children and Department of Molecular and Medical Genetics University of Toronto, 555 University Ave., Toronto , M5G 1X8, ON, Canada

    Guillermo Guenechea, Olga I. Gan, Craig Dorrell & John E. Dick

  2. Gene Therapy Program, CIEMAT/Fundación Marcelino Botín, Madrid, Spain

    Guillermo Guenechea

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  1. Guillermo Guenechea
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Correspondence to John E. Dick.

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Guenechea, G., Gan, O., Dorrell, C. et al. Distinct classes of human stem cells that differ in proliferative and self-renewal potential. Nat Immunol 2, 75–82 (2001). https://doi.org/10.1038/83199

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