Hoang T, Gilmore D, Metcalf D, Cobbold S, Watt S, Clark M, Furth M, Waldmann H
Blood. 1983 Mar;61(3):580-8.
Primitive hemopoietic progenitor cells from adult mouse marrow have been substantially enriched by virtue of a negative selection procedure with monoclonal antibodies. It has been possible to segregate erythroid progenitor cells at distinct stages of differentiation on the basis of their cell surface antigens. This has been achieved with two monoclonal antibodies reactive with the mature elements of bone marrow. YBM 34.3 binds to a heat-stable antigen expressed on B lymphocytes, neutrophils, and cells of the erythroid lineage. YBM 6.1 reacts with cells of the neutrophil, eosinophil, and monocyte series but does not bind to colony-forming cells. Separation is achieved by indirect immunoadsorption (panning) with YBM 34.3 on Protein-A-coated plastic plates followed by FACS II cell sorting with YBM 6.1. The combined procedures yield a marrow population containing 58% immature cells (blasts, promyelocytes, and myelocytes) and 9.5% clonogenic cells. In addition, differential binding of YBM 34.3 can be used to segregate erythroid progenitor cells at distinct stages of differentiation (day 7 BFU-E, day 5 BFU-E and CFU-E) either by cell sorting or panning. It is shown that both techniques give a comparable degree of resolution of the different cell types with, however, an appreciable advantage of panning over cell sorting in allowing the rapid handling of large numbers of cells.
通过用单克隆抗体进行阴性选择程序，已从成年小鼠骨髓中大量富集了原始造血祖细胞。基于其细胞表面抗原，已能够在不同分化阶段分离红系祖细胞。这是通过两种与骨髓成熟成分反应的单克隆抗体实现的。YBM 34.3与B淋巴细胞、中性粒细胞和红系谱系细胞上表达的一种热稳定抗原结合。YBM 6.1与中性粒细胞、嗜酸性粒细胞和单核细胞系列的细胞反应，但不与集落形成细胞结合。通过在蛋白A包被的塑料板上用YBM 34.3进行间接免疫吸附（淘选），然后用YBM 6.1进行FACS II细胞分选来实现分离。联合程序产生的骨髓群体含有58%的未成熟细胞（原始细胞、早幼粒细胞和中幼粒细胞）和9.5%的克隆形成细胞。此外，YBM 34.3的差异结合可用于通过细胞分选或淘选在不同分化阶段（第7天的爆式红系集落形成单位、第5天的爆式红系集落形成单位和红系集落形成单位）分离红系祖细胞。结果表明，两种技术对不同细胞类型的分辨程度相当，然而，淘选在允许快速处理大量细胞方面比细胞分选具有明显优势。
