Somatic cell hybridisation

This is a technique pivotal to the genetic analysis of cultured animal cells and to the production of monoclonal antibodies (see 13.6) (Ringertz and Savage, 1976 Campbell, 1984). 

Although such studies are crude relative to the genetic analysis in bacteria, it is now easy to locate genes to particular chromosomes. Traditionally, such studies have been performed by analysis of the offspring of parents showing particular phenotypic characteristics, but the process was taken to the level of biochemical characteristics 

In this way it has been shown that HPRT is located on the X-chromosome and much work in the 1970 s developed from such observations (Goss and Harris, 1975 Willecke et al., 1976a, b). 

Human cells are killed by 10 7 ouabain, but mouse cells are relatively resistant in that 10 3M is a lethal dose. Mouse human hybrids show intermediate sensitivity and hence HAT medium containing 10 7M ouabain can be used to select for hybrids between any human cell and TK or HPRT mouse cells. This enables primarily human cells and other non-selected strains to be used in fusion experiments (Mayhew, 1972 Thompson and Baker, 1973). 

Other selection systems involve the fusion of proline requiring CHO cells and defective mouse cells in HAT medium lacking proline and the fusion of normal lymphocytes (which do not grow in vitro) with defective mouse cells (which do not grow in HAT medium). It is this latter technique which has allowed the isolation of clones of cells which will synthesise in vitro large amounts of a single antibody (i.e. a monoclonal antibody) (Kohler, 1982). 

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Ephrussi, B. (1972). Hybridisation of Somatic Cells (New Jersey Princeton University Press)... 

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