Jurecic R, Van N T, Belmont J W
Institute for Molecular Genetics, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030.
Blood. 1993 Nov 1;82(9):2673-83.
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.
来自FVB/N和BALB/c小鼠（Ly-6a单倍型）的所有骨髓（BM）细胞中约4%至5%以及低密度BM细胞中8%至9%显示出Ly-6E.1抗原的高至中等表达，该抗原可被Sca-1抗体识别。对表达Sca-1抗原的Ly-6E.1等位基因形式的富集细胞的功能特性进行了分析，并与表达凝集素麦胚凝集素（WGA）碳水化合物结合位点的细胞特性相关联。使用平衡密度离心和荧光激活细胞分选技术，分离出Sca-1+WGA+、Lin-WGA+、Lin-Sca-1+和Lin-Sca-1+WGA+细胞，并测定了它们的脾集落形成单位（CFU-S）细胞含量、辐射防护能力和长期重建能力。富集的Sca-1+WGA+、Lin-WGA+、Lin-Sca-1+和Lin-Sca-1+WGA+细胞分别在26、20、21和15个分选细胞中产生1个CFU-S12细胞。当移植到致死性照射的受体（100至500个细胞/小鼠）中时，在30天的辐射防护试验中，所有群体拯救了70%至100%的受体，并在移植后6个月介导了40%至80%的受体存活。使用转基因小鼠作为细胞供体，我们已经表明，在移植100个Sca-1+WGA+、Lin-WGA+、Lin-Sca-1+和Lin-Sca-1+WGA+细胞12至16周后，40%至80%的受体在骨髓、脾脏、胸腺和淋巴结中有供体细胞。这些结果表明，在两种分析的具有Ly-6a单倍型的小鼠品系中，表达Sca-1抗原的Ly-6E.1形式的细胞群体包含CFU-S和长期重建细胞。此外，数据表明，至少在FVB/N小鼠中，第12天的CFU-S细胞和具有长期重建能力的细胞同时表达Sca-1抗原的Ly-6E.1形式和WGA结合分子。
