Abstract
Ts systems provide an opportunity to induce a cell with a defined genotype to express either the transformed or normal phenotype by simply manipulating temperature. Rat kidney fibroblasts transformed by a Ts derivative of the Kirsten murine sarcoma virus grow on agar or a preexisting monolayer at the permissive temperature (32°C) but are unable to grow under these conditions at the nonpermissive temperature (39°C). The purpose of this study was to determine the suitability of this model to study processes accompanying “phenotypic transformation” (normal ⇌ transformed). Growth characteristics (clonal/mass culture), cell size, cAMP levels, and surface morphology were periodically monitored following the manipulation of temperature from 39°C ⇌ 32°C. Ts, normal and transformed control cells grew slower at 32°C, but attained a saturation density equivalent to cells grown at 39°C. Cell size was inversely related to the growth rate; each cell type was smallest during the period of maximal growth rate and became larger as division slowed and cells reached confluency. cAMP levels were also inversely related to growth rate; all cells growing at similar rates had similar cAMP levels. Scanning EM showed that cells maintained at 32°C became spindle-shaped and “piled up” at low density and low serum concentration while cells at 39°C formed flat, confluent monolayers and required greater serum concentration for growth. This system may provide the means to distinguish cellular changes due to transformation from those common to “normal” cells with similar growth rates.
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Guinivan, P., Ladda, R. AN IN VITRO MODEL FOR THE STUDY OF TRANSFORMATION: TEMPERATURE-SENSITIVE (TS) MUTANT. Pediatr Res 11, 457 (1977). https://doi.org/10.1203/00006450-197704000-00522
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DOI: https://doi.org/10.1203/00006450-197704000-00522