van den Heuvel M, Wilson J, McBey B A, Yamashiro S, Croy B A
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Canada.
J Reprod Immunol. 1994 Aug;27(1):13-36. doi: 10.1016/0165-0378(94)90012-4.
Seven independent cell lines were derived from the fusion of migratory cells recovered from explant cultures of metrial glands to SP 2/0, a non-Ig secreting B cell myeloma. The migrating cells came from a pool of metrial glands from day 6-8 pregnant random bred CD1 mice and were assumed to be cells early in the differentiation pathway to granulated metrial gland (GMG) cells. The fused cells were cloned twice at the limiting dilution. Hybridization was confirmed by quantitation of cellular DNA using propidium iodide staining and by karyotyping. Electron microscopy revealed that each of the hybrid cell lines was composed of cells which were lymphoid in appearance, but lacked the granules found in mature GMG cells. The surface phenotype of all lines is CD45+, LGL-1-, asialo GM-1-, IgG-, IgM-, CD3- and CD25- (p55 of IL-2 receptor). Although the hybridomas lack those phenotypic markers which were used to show that GMG cells are related to the natural killer (NK) cell lineage (ie LGL-1, asialo GM-1), they do express the pan-leukocyte marker CD45 as well as the lytic protein, perforin, at levels intermediate to those of SP 2/0 cells and GMG cells. In addition, the hybridomas were observed to preferentially bind the NK target cell YAC and to be capable of lytic activity at temperatures below 30 degrees C. Because these hybridomas may represent fusion to an early progenitor cell of the NK/GMG cell lineage, their continued characterization is of merit.
从子宫内膜腺外植体培养物中回收的迁移细胞与SP 2/0(一种非Ig分泌性B细胞骨髓瘤)融合,获得了7个独立的细胞系。迁移细胞来自妊娠第6 - 8天的随机繁殖CD1小鼠的一组子宫内膜腺,被认为是向颗粒化子宫内膜腺(GMG)细胞分化途径早期的细胞。融合细胞在极限稀释条件下克隆了两次。通过使用碘化丙啶染色定量细胞DNA和核型分析证实了杂交。电子显微镜显示,每个杂交细胞系均由外观呈淋巴细胞样但缺乏成熟GMG细胞中发现的颗粒的细胞组成。所有细胞系的表面表型均为CD45 +、LGL - 1 -、去唾液酸GM - 1 -、IgG -、IgM -、CD3 -和CD25 -(IL - 2受体的p55)。尽管杂交瘤缺乏那些用于表明GMG细胞与自然杀伤(NK)细胞谱系相关的表型标记(即LGL - 1、去唾液酸GM - 1),但它们确实表达泛白细胞标记CD45以及溶细胞蛋白穿孔素,其表达水平介于SP 2/0细胞和GMG细胞之间。此外,观察到杂交瘤优先结合NK靶细胞YAC,并且在低于30摄氏度的温度下具有溶细胞活性。由于这些杂交瘤可能代表与NK/GMG细胞谱系的早期祖细胞融合,因此对它们的持续表征具有重要意义。