Enrichment and Functional Characterization of Sca-1 + WGA +, Lin WGA +, Lin− Sca-1+, and Lin− Sea-1+ WGA+ Bone Marrow Cells From Mice With an Ly-6a Haplotype

Abstract

AbstractApproximately 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, ra-dioprotection 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.