Zheng Y, McNeill K, Rector E S, Froese A
Department of Immunology, University of Manitoba, Winnipeg, Canada.
Int Arch Allergy Immunol. 1995 Nov;108(3):231-9. doi: 10.1159/000237158.
Rat peritoneal mast cells and 6-thioguanine-resistant rat basophilic leukemia cells, representative of connective tissue-type (CTMC) and mucosal (MMC) mast cells, respectively, were fused using polyethylene glycol. Four out of 14 primary hybrid mast cell lines contained more than 50% of CTMC as demonstrated by histochemical staining. Two cell lines, one predominantly of the CTMC and the other of the MMC phenotype, were selected for further study. Among these, the phenotype was also confirmed by analysis for rat mast cell protease I and by mediator release triggered by compound 48/80 and ionophore A23187. The CTMC phenotype disappeared after culturing cells for 2 weeks. The change in phenotype did not significantly alter the mediator release due to calcium ionophore A23187. Repeated cloning of cells bearing the CTMC phenotype did not yield a cloned line of cells expressing the CTMC phenotype only, although it prolonged the persistence of this phenotype. During the period of CTMC phenotype loss, a drop in cellular DNA content occurred, suggesting that chromosome instability may, at least partially, have been responsible for the phenotypic changes.
分别代表结缔组织型(CTMC)和黏膜型(MMC)肥大细胞的大鼠腹膜肥大细胞和耐6-硫鸟嘌呤大鼠嗜碱性白血病细胞,使用聚乙二醇进行融合。通过组织化学染色证明,14个初代杂交肥大细胞系中有4个含有超过50%的CTMC。选择了两个细胞系,一个主要为CTMC表型,另一个为MMC表型,进行进一步研究。其中,通过对大鼠肥大细胞蛋白酶I的分析以及由化合物48/80和离子载体A23187触发的介质释放,也证实了表型。在培养细胞2周后,CTMC表型消失。表型的变化并未显著改变由钙离子载体A23187引起的介质释放。对具有CTMC表型的细胞进行反复克隆,并未产生仅表达CTMC表型的克隆细胞系,尽管这延长了该表型的持续时间。在CTMC表型丧失期间,细胞DNA含量下降,表明染色体不稳定性可能至少部分地导致了表型变化。