Suppression of transformation and immortality in human/Chinese hamster fibroblast hybrids--a model for suppressor gene isolation.
Somatic cell hybrids were produced by fusion of normal human (foreskin) fibroblasts and a transformed Chinese hamster fibroblast line V79-8. Overall, approximately 30% of hybrid clones showed stable reversion to normal morphology and growth control in vitro as shown by serum and anchorage dependence. In one-third of these clones, senescence was observed after a number of generations similar to that required for the human fibroblast parent cells to senesce. The remainder appear to be immortal. Normal human chromosomes can therefore restore growth control with or without finite life-span to this transformed cell. V79 cells were found to be transfectable at an efficiency compatible with detection of single-copy gene transfer from genomic DNA. Furthermore, these cells were exceptionally sensitive to negative ("suicide") selection. Taken together, our data suggest that the V79 line represents an ideal system for isolation of human tumour suppressor genes.
Wynford-Thomas
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Department of Pathology, University of Wales College of Medicine, Cardiff, UK.
Bond
J A
JA
Paterson
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D000818:Animals; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000187:drug effects; D007621:Karyotyping; D010641:Phenotype; D011092:Polyethylene Glycols / Q000494:pharmacology; D013489:Suppression, Genetic; D013997:Time Factors; D014162:Transfection
D011092:Polyethylene Glycols; D004247:DNA