A newly discovered class of human hematopoietic cells with SCID-repopulating activity

Abstract

The detection of primitive hematopoietic cells based on repopulation of immune-deficient mice is a powerful tool to characterize the human stem-cell compartment. Here, we identify a newly discovered human repopulating cell, distinct from previously identified repopulating cells, that initiates multilineage hematopoiesis in NOD/SCID mice. We call such cells CD34neg- S CID r epopulating c ells, or CD34neg-SRC. CD34neg-SRC are restricted to a LinCD34CD38 population without detectable surface markers for multiple lineages and CD38 or those previously associated with stem cells (HLA-DR, Thy-1 and CD34). In contrast to CD34+ subfractions, LinCD34CD38 cells have low clonogenicity in short-and long-term in vitro assays. The number of CD34neg-SRC increased in short-term suspension cultures in conditions that did not maintain SRC derived from CD34+ populations, providing independent biological evidence of their distinctiveness. The identification of this newly discovered cell demonstrates complexity of the organization of the human stem-cell compartment and has important implications for clinical applications involving stem-cell transplantation.

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Figure 1: Phenotypic analysis of human LinCD34 hematopoietic cells.
Figure 2: Frequency analysis of LinCD34 cells found in human hematopoietic tissue throughout development.
Figure 3: Human cell engraftment of NOD/SCID mice transplanted at various cell doses with purified LinCD34 cells from cord blood.
Figure 4: Multilineage differentiation of human LinCD34 cells in NOD/SCID mice.
Figure 5: Capacity of LinCD34, LinCD34CD38, LinCD34CD38+ and LinCD34+CD38 cells cell fractions to engraft NOD/SCID after ex vivo culture.

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Acknowledgements

We thank L. McWhirter for providing cord blood and fetal specimens, H. Messner and N. Jamal for providing bone marrow and peripheral blood samples, G. Knowles for technical support in the flow cytometric analysis, and R. Phillips, A. Bernstein, N. Iscove, R. McInnes and members of the laboratory for reviewing the manuscript. This work was supported by grants to J.E.D. from the Medical Research Council of Canada (MRC), the National Cancer Institute of Canada (NCIC) with funds from the Canadian Cancer Society, the Canadian Genetic Diseases Network of the National Centers of Excellence, an MRC Scientist award (J.E.D.), postdoctoral fellowships from the NCIC (M.B.), and the Human Frontier Science Organization Program (D.B.).

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Correspondence to John E. Dick.

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Bhatia, M., Bonnet, D., Murdoch, B. et al. A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat Med 4, 1038–1045 (1998) doi:10.1038/2023

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