Growth in high serum concentrations leads to rapid deadaptation of cells previously adapted to growth in an extremely low concentration of serum.

A subline of NIH 3T3 cells adapted to multiply in 0.25% calf serum (CS) by frequent passage (every 2-3 days) at low population density in 0.25% CS was deadapted by frequent successive passages of the cells in 10% CS for 3 weeks. The cells adapted to 0.25% CS multiplied with an average doubling time of 16.9 hr in 10% CS, and cells that had always been kept in 10% CS multiplied with an average doubling time of 14.6 hr, so there was weak selection for the latter in the higher serum concentration. When adapted cells were subjected to two passagers in 10% CS prior to assay of growth in 0.25% CS, a 4-day lag period was evident before commencement of exponential growth, and there was a decrease in saturation density. Further delay of growth in 0.25% CS developed as the number of passages of cells in 10% CS increased. The marked delay of growth in 0.25% CS of the bulk population after a few days in 10% CS argued against selection in 10% CS of rare nonadapted mutants from the adapted population and for an epigenetic origin of the change. Reconstruction experiments utilizing adapted cells mixed with non-adapted cells in 0.25% CS buttressed this explanation. Eight clones of the adapted population exhibited some loss of growth capacity in 0.25% CS after a single passage in 10% CS, though the extent of loss varied from clone to clone. The results support the idea that all cells in the adapted population respond to the lifting of growth constraints with loss of their growth potential under highly constrained conditions. They are consistent with the concept of progressive state selection in which selection operates on fluctuating metabolic states of individual cells rather than on genetic variants.