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Blood, 15 August 2000, Vol. 96, No. 4, pp. 1380-1387
HEMATOPOIESIS
Homing and engraftment potential of
Sca-1+lin cells fractionated on the basis of
adhesion molecule expression and position in cell cycle
Christie M. Orschell-Traycoff,
Kelly Hiatt,
Ramzi N. Dagher,
Susan Rice,
Mervin C. Yoder, and
Edward F. Srour
From the Division of Hematology/Oncology and Indiana
Elks Cancer Research Center, Department of Medicine; Herman B Wells
Center for Pediatric Research, Department of Pediatrics; and Department
of Microbiology and Immunology, Indiana University School of Medicine,
Indianapolis.
Engraftment potential of hematopoietic stem cells (HSCs) is likely
to be dependent on several factors including expression of
certain adhesion molecules (AMs) and degree of mitotic quiescence. The
authors investigated the functional properties and engraftment potential of Sca-1+lin cells subfractionated
on the basis of expression, or lack thereof, of CD11a, CD43, CD49d,
CD49e, or CD62L and correlated that expression with cell cycle
status and proliferative potential of engrafting fractions.
Donor-derived chimerism in mice receiving CD49e+ or
CD43+ Sca-1+lin cells was greater
than that in mice receiving cells lacking these 2 markers, while
Sca-1+lin cells positive for CD11a and CD62L
and bright for CD49d expression mediated minimal engraftment. AM
phenotypes enriched for engraftment potential contained the majority of
high proliferative potential-colony forming cells, low
proliferative potential-colony forming cells, and cells providing
rapid in vitro expansion. Cell cycle analysis of AM subpopulations
revealed that, regardless of their bone marrow repopulating potential,
Sca-1+lin AM cells contained a
higher percentage of cells in G0/G1 than their AM+ counterparts. Interestingly, engrafting phenotypes,
regardless of the status of their AM expression, were quicker to exit
G0/G1 following in vitro cytokine stimulation
than their opposing phenotypes. When engrafting phenotypes of
Sca-1+lin AM+ or AM
cells were further fractionated by Hoechst 33342 into
G0/G1 or S/G2+M, cells providing
long-term engraftment were predominantly contained within the quiescent
fraction. These results define a theoretical phenotype of a
Sca-1+lin engrafting cell as one that is
mitotically quiescent, CD43+, CD49e+,
CD11a, CD49ddim, and CD62L.
Furthermore, these data suggest that kinetics of in vitro proliferation may be a good predictor of engraftment potential of candidate populations of HSCs.
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