Abstract
The fusion of different types of cultured mammalian cells to produce hybrid cells and heterokaryons is now an established research tool that is widely used in many areas of cell biology and virology. The use of viruses, Sendai virus in particular, to fuse cells was a major factor in the successful development of cell-fusion techniques in the 1960s, and viruses remain the most commonly used agents for inducing cell fusion. Although viruses are undoubtedly highly efficient agents for inducing cell fusion in vitro , there are certain practical difficulties associated with their use. First, the virus components responsible for inducing cell fusion are not fully characterized, and therefore the fusion potential of different virus preparations cannot be standardized. The use of lysolecithin and similar lipolytic agents to induce cell fusion is accompanied by substantial cell damage, and the number of viable cells recovered is significantly lower than in cells treated with viruses. Consequently, there is still a need for a simple procedure for inducing cell fusion that avoids the problems accompanying fusion induced by inactivated viruses or membrane lytic agents, such as lysolecithin. Potential progress in this direction has been made with the recent demonstration that small vesicles (approximately 250-500 A diameter) prepared from a variety of phospholipids are able to induce fusion of mammalian cells in vitro. Lipid vesicles compare favorably with inactivated Sendai virus in their ability to fuse cells, yet are devoid of the cytotoxic properties associated with lysolecithin. This chapter presents an outline of the methods used in this laboratory to produce lipid vesicles for inducing fusion and hybridization of cultured mammalian cells.
Published Version
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