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Enrichment and functional characterization of Sca-1+WGA+, Lin-WGA+, Lin-
Sca-1+, and Lin-Sca-1+WGA+ bone marrow cells from mice with an Ly-6a
haplotype
R Jurecic, NT Van and JW Belmont
Institute for Molecular Genetics, Howard Hughes Medical Institute, Baylor
College of Medicine, Houston, TX 77030.
Approximately 4% to 5% of all bone marrow (BM) cells and 8% to 9% of low
density BM cells from FVB/N and BALB/c mice (Ly-6a haplotype) show high to
intermediate expression of Ly-6E.1 antigen, recognized by the Sca-1
antibody. Functional properties of enriched cells expressing Ly-
6E.1-allelic form of Sca-1 antigen were analyzed and correlated with the
properties of cells expressing the carbohydrate binding sites for the
lectin wheat-germ agglutinin (WGA). Using equilibrium density
centrifugation and fluorescence-activated cell sorting, Sca-1+WGA+, Lin-
WGA+, Lin-Sca-1+, and Lin-Sca-1+WGA+ cells were isolated and their splenic
colony-forming unit (CFU-S) cell content, radioprotection ability, and
long-term reconstitution capacity determined. Enriched Sca- 1+WGA+,
Lin-WGA+, Lin-Sca-1+ and Lin-Sca-1+WGA+ cells gave rise to 1 CFU-S12 cell
out of 26, 20, 21, and 15 sorted cells, respectively. When transplanted
into lethally irradiated recipients (100 to 500 cells/mouse) all
populations rescued 70% to 100% of recipients in a 30- day radioprotection
assay and mediated survival of 40% to 80% of recipients 6 months after
transplantation. Using transgenic mice as cell donors we have shown that 12
to 16 weeks after transplantation of 100 Sca-1+WGA+, Lin-WGA+, Lin-Sca-1+,
and Lin-Sca-1+WGA+ cells, 40% to 80% of recipients had donor cells in BM,
spleen, thymus, and lymph nodes. These results indicate that the population
of cells expressing Ly-6E.1 form of Sca-1 antigen in two analyzed mouse
strains with Ly-6a haplotype contains CFU-S and long-term repopulating
cells. Furthermore, the data suggest that, at least in FVB/N mice, day-12
CFU-S cells and cells with long-term repopulating capacity simultaneously
express Ly- 6E.1 form of Sca-1 antigen and WGA-binding molecules.
Volume 82,
Issue 9,
pp. 2673-2683,
11/01/1993
Copyright © 1993 by The American Society of Hematology
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