Radomska H S, Eckhardt L A
Department of Biological Sciences, Hunter College, City University of New York, NY 10021, USA.
J Immunol Methods. 1995 Dec 27;188(2):209-17. doi: 10.1016/0022-1759(95)00205-7.
We and others have been interested in the phenomenon of gene 'extinction' in somatic cell hybrids, reasoning that the study of this process is likely to reveal underlying mechanisms responsible for limiting the expression of specialized genes only to appropriate cell types. In the course of our studies in this area, we have developed a simple and economical method of fusing mammalian cells, using an electroporation device. In fusions between murine myeloma and T lymphoma lines, hybrid cell recoveries were typically one per 10(5) [corrected] input myeloma cells. Because of our interest in the regulation of immunoglobulin heavy chain (IgH) gene expression, we analyzed the hybrids for both IgH gene composition and expression. The hybrid lines were phenotypically indistinguishable from those generated by the more conventional, polyethylene glycol (PEG)-induced fusion protocol. There was a notable increase, however, in the number of hybrids that retained IgH-encoding chromosomes from both parental lines.
我们和其他研究人员一直对体细胞杂种中基因“灭绝”现象感兴趣,理由是对这一过程的研究可能会揭示将特定基因的表达限制在适当细胞类型的潜在机制。在我们对该领域的研究过程中,我们开发了一种使用电穿孔装置融合哺乳动物细胞的简单且经济的方法。在小鼠骨髓瘤细胞系与T淋巴瘤细胞系的融合中,杂种细胞回收率通常为每10⁵个[校正后]输入的骨髓瘤细胞产生1个。由于我们对免疫球蛋白重链(IgH)基因表达的调控感兴趣,我们分析了杂种细胞的IgH基因组成和表达情况。这些杂种细胞系在表型上与通过更传统的聚乙二醇(PEG)诱导融合方案产生的细胞系没有区别。然而,保留来自两个亲代细胞系的IgH编码染色体的杂种细胞数量显著增加。