ELECTROFUSION

This chapter provides an overview of electrofusion. Electrofusion offers several advantages over conventional methods including the ability to produce hybridomas with much smaller numbers of fusion partners, higher fusion efficiency, higher hybrid viability, effectiveness with a wider variety of cell types, and a more controlled, reproducible approach. This chapter discusses the developments and outlines current electrofusion technology and protocols. Successful application of the electrofusion technique requires an understanding of the rudiments of cell membrane behavior in electrical fields. The fundamental step in electrofusion is reversible membrane breakdown. When short-duration electrical impulses applied across a cell membrane exceed a critical threshold, that membrane becomes transiently, but highly permeable. The electrofusion medium is of critical importance for cell viability and successful hybridoma production. To minimize potentially deleterious manipulations and shock to the cells, a single medium is preferred for alignment, fusion, and resealing. © 1993, Elsevier Inc. All rights reserved.