Craig W H, Thomas T E, Lansdorp P M
Terry Fox Laboratory, B.C. Cancer Research Centre, Vancouver, Canada.
J Hematother. 1992 Spring;1(1):55-64. doi: 10.1089/scd.1.1992.1.55.
The binding of lectins to nucleated cells from human bone marrow was studied in a search for markers that can be used to subdivide further immature hemopoietic cells that are characterized by their expression of CD34. Low-density bone marrow cells were indirectly labeled with biotinylated lectins and streptavidin-R-phycoerythrin (SA-RPE) together with FITC-labeled monoclonal anti-CD34. Four-parameter flow cytometric analysis was then performed and list mode data analyzed. Of the 21 lectins tested, only a few showed differential staining of CD34+ versus CD34- cells. These include soybean agglutinin (SBA) and Ulex europaeus agglutinin I (UE). Lycopersicon esculentum (LE) and Erythrina cristigalli (EC) reacted preferentially with, respectively, CD34+ and CD34- cells, suggesting their usefulness in some method to enrich for CD34+ cells. This possibility was tested by passing cells labeled with biotinylated lectins over a column containing streptavidin-coated beads. CD34+ cells could be enriched > 10-fold by competitive (sugar) elution of LE-labeled cells from the column. Similarly, depletion of biotinylated EC-labeled cells by passage through the streptavidin column enriched CD34+ cells several fold. The results of these studies document the reactivity of a large panel of lectins with subpopulations of nucleated bone marrow cells and indicate that certain lectins could possibly be used for development of cell separation procedures aimed at the selective enrichment of cells that express CD34.
为寻找可用于进一步细分以表达CD34为特征的未成熟造血细胞的标志物，研究了凝集素与人骨髓有核细胞的结合情况。用生物素化凝集素和链霉亲和素 - R - 藻红蛋白（SA - RPE）间接标记低密度骨髓细胞，并同时使用异硫氰酸荧光素（FITC）标记的抗CD34单克隆抗体。然后进行四参数流式细胞术分析并分析列表模式数据。在所测试的21种凝集素中，只有少数几种对CD34 +细胞和CD34 -细胞显示出差异染色。其中包括大豆凝集素（SBA）和欧洲荆豆凝集素I（UEA）。番茄凝集素（LE）和刺桐凝集素（EC）分别优先与CD34 +细胞和CD34 -细胞反应，表明它们在某种富集CD34 +细胞的方法中有用。通过将用生物素化凝集素标记的细胞通过含有链霉亲和素包被珠子的柱子来测试这种可能性。通过从柱子上竞争性（糖）洗脱LE标记的细胞，CD34 +细胞可以富集超过10倍。同样，通过链霉亲和素柱使生物素化EC标记的细胞耗尽，可使CD34 +细胞富集几倍。这些研究结果证明了一大组凝集素与有核骨髓细胞亚群的反应性，并表明某些凝集素可能用于开发旨在选择性富集表达CD34细胞的细胞分离程序。
