Abstract
This chapter describes how somatic cell hybridization has not only shown that tissue-specific genes are regulated by transacting factors, but also has provided strong evidence for the existence of tumor suppressor gene frequency. A spontaneous fusion of cells in culture occurs at a very low frequency. The best known of such methods is the application of hypoxanthine + aminopterin + thymidine (HAT) selection for the fusion of cells deficient in hypoxanthine guanosine phosphoryl transferase with cells deficient in thymidine kinase, but different combinations of selectable markers can be used. Inoculate the mixture of parental cells to be fused into several 50-ram tissue culture dishes containing complete growth medium. The total number of cells has to be adjusted to occupy the entire dish surface, such that the fusion will be done on cells that are in close contact. Resuspend thoroughly the cell pellet in a complete growth medium and count the cell number in a hemacytometer.
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