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Related Experiment Videos

Long-term hemopoietic repopulation by Thy-1lo, Lin-, Ly6A/E+ cells.

C L Li1, G R Johnson

  • 1Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.

Experimental Hematology
|December 1, 1992
PubMed
Summary

Hematopoietic stem cell populations containing Thy-1lo, Lin-, Ly6A/E+ cells can reconstitute myeloid and lymphoid lineages in mice. This cell population is heterogeneous, containing both long-term repopulating cells and lymphoid-restricted progenitors.

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Area of Science:

  • Hematology
  • Immunology
  • Stem Cell Biology

Background:

  • Hematopoietic stem cells (HSCs) are crucial for maintaining blood cell production.
  • Identifying specific HSC populations is key to understanding hematopoiesis and developing cell therapies.
  • Murine bone marrow Thy-1lo, Lin-, Ly6A/E+ cells represent a potential HSC population.

Purpose of the Study:

  • To characterize the repopulating capacity of murine bone marrow Thy-1lo, Lin-, Ly6A/E+ cells.
  • To determine the lineage potential (myeloid and lymphoid) of these cells.
  • To assess the heterogeneity within this cell population.

Main Methods:

  • Isolation of murine bone marrow Thy-1lo, Lin-, Ly6A/E+ cells using fluorescence-activated cell sorting (FACS).
  • In vitro colony-forming cell (CFC) assays stimulated with interleukin-3 (IL-3).

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  • In vivo transplantation into lethally irradiated recipients to assess spleen colony-forming units (CFU-S) and long-term reconstitution.
  • Analysis of peripheral blood cell populations in primary and secondary recipients over 72 weeks.

    • Main Results:

    • The Thy-1lo, Lin-, Ly6A/E+ population contained up to 27% in vitro CFC and 5% day-12 CFU-S.
    • As few as 100 cells could reconstitute myeloid and lymphoid compartments for up to 72 weeks.
    • Engraftment patterns varied, with some recipients showing sustained high proportions of donor cells (>90%), while others showed decline or static levels.
    • High donor cell percentages (>60%) in peripheral blood indicated multilineage reconstitution (myeloid and lymphoid).
    • Lower donor cell percentages (<20%) primarily led to lymphoid reconstitution in secondary recipients.
    • The cell population demonstrated heterogeneity, with some cells capable of long-term myeloid and lymphoid repopulation, and others restricted to lymphoid lineage.

    Conclusions:

    • The Thy-1lo, Lin-, Ly6A/E+ murine bone marrow cell population is heterogeneous.
    • It contains cells with potent in vitro and in vivo hematopoietic stem cell activity, including long-term repopulating cells for both myeloid and lymphoid lineages.
    • The data suggest the presence of both multipotent and lymphoid-restricted progenitors within this sorted population.