Harvesting of myeloma cells

Myeloma cells (e.g. the rat Y3.Ag.l.2.3 cell line—Galfre et al., 1979) must be growing exponentially when harvested. They should be pelletted at 800 g and washed in serum-free medium. 

2-5 X 107 myeloma cells are required to fuse to 10s spleen cells and fusion techniques are described in 13.7. 

Although cells sometimes fuse spontaneously, the frequency of fusion can be dramatically increased by treatment with a number of reagents. Sendai virus used to be a very common fusogen (Watkins, 1971 Stadler and Adelbert, 1972) but is difficult to control. It has been largely replaced by lysolecithin or polyethyleneglycol (PEG) or by electrofusion. 

This method devised by Pontecorvo (Davidson et al., 1976 Ponte-corvo et al., 1977) is the simplest method and the one in most 

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