Evidence for the progressive and adaptive nature of spontaneous transformation in the NIH 3T3 cell line.

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Abstract
The NIH 3T3 mouse cell line is widely used as a recipient of DNA from tumors to demonstrate the presence of transforming oncogenes. We show that these cells produce transformed foci spontaneously if kept in the confluent state for more than 10 days. The formation of foci depends on the type and concentration of bovine serum used in the medium and passage history of the cells. Cells maintained in continuous exponential multiplication in the subconfluent state by transfer every 2-3 days in medium with 10% calf serum failed to develop the capacity to produce foci in 2% calf serum, but those transferred the same way in 2% calf serum or in 10% fetal bovine serum, which is a less potent growth stimulant, did develop that capacity to an increasing degree over time. The number of transformed cells increased sharply with the time that a culture remained in the confluent state. There are several morphological types and degrees of transformation, which indicates that the underlying changes were varied and the process is progressive. The results also suggest that transformation occurs in a fraction of an entire cell population that is undergoing a physiological adaptation to moderate constraints on its growth.