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细胞杂交与细胞凝集。II. 聚阳离子对细胞杂交的促进作用。

Cell hybridization and cell agglutination. II. Enhancement of cell hybridization by polycations.

作者信息

Matsuya Y, Yamane I

出版信息

J Cell Sci. 1985 Oct;78:273-82. doi: 10.1242/jcs.78.1.273.

Abstract

An efficient technique for hybridization of mammalian cells was developed by combining agglutination by pretreatment with polycations, such as polyarginine, and conventional polyethylene glycol(PEG)-mediated cell fusion. Polyarginine and subsequent PEG treatment resulted in markedly decreased viability in the treated cells, but addition of polyvinyl pyrrolidone or glycerol to the polyarginine prevented this cytotoxicity. Polyarginine was much more effective than polylysine or polyornithine in inducing hybridization. Other polycations, including polybrene and protamine but not DEAE-dextran, were also active in inducing hybridization. The condition of the cells at the time of polycation treatment was an important factor in the enhancement of hybridization. The condition of the cells at the time of polycation treatment was an important factor in the enhancement of hybridization. The enhancement of hybridization of cells in monolayer incubated for 2 h was much higher than that of cells incubated for 24 h. These findings suggest that polycations do not necessarily operate by agglutinating cells. The mechanism of polycation-enhanced cell hybridization is discussed.

摘要

通过将用聚阳离子(如聚精氨酸)预处理的凝集作用与传统的聚乙二醇(PEG)介导的细胞融合相结合,开发出了一种用于哺乳动物细胞杂交的高效技术。聚精氨酸和随后的PEG处理导致处理过的细胞活力显著降低,但在聚精氨酸中添加聚乙烯吡咯烷酮或甘油可防止这种细胞毒性。在诱导杂交方面,聚精氨酸比聚赖氨酸或聚鸟氨酸有效得多。包括聚凝胺和鱼精蛋白但不包括DEAE-葡聚糖在内的其他聚阳离子在诱导杂交方面也有活性。聚阳离子处理时细胞的状态是增强杂交的一个重要因素。单层培养2小时的细胞杂交增强程度远高于培养24小时的细胞。这些发现表明聚阳离子不一定通过凝集细胞起作用。文中讨论了聚阳离子增强细胞杂交的机制。

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