The 3T3 cell cycle at low proliferation rates.

When the proliferation rate of Swiss 3T3 cells is decreased by limiting the availability of growth factors, cell cycle variability increases, as predicted by the transition probability model. Nevertheless, the transition probabilities would appear to play a relatively minor role in the regulation of proliferation rate. Instead, at least 40% of the increase in the average cycle time is brought about by an elongation of the minimum cycle time (i.e. the 'deterministic' part of the cycle). In addition, we have found that a substantial proportion of the cells (roughly 20%, in the present experiments, for doubling times of the order of 35-40 h) drop out of cycle in each generation, leading to a growth fraction of less than 1.0. The non-dividing cells, which we have previously shown to remain capable of division, would seem to support the existence of a Go state outside the normal cell cycle, and distinct from the indeterminate states postulated by the transition probability model. Because of the generation of nondividing cells at low proliferation rates, the log alpha and beta plots (distributions of cycle times, and sibling cycle time differences, respectively) are markedly concave, with a continuously decreasing slope. The transition probabilities cannot therefore be estimated directly and it is impossible to determine the extent to which they contribute to the regulation of proliferation rate. Rather, our data suggest that the transition probabilities are not uniform throughout the population under these conditions, but vary substantially from cell to cell. In addition to the changes in cell cycle kinetics, we also report an increased failure rate of cytokinesis, at low proliferation rates, leading initially to the appearance of binucleate cells. Such failures of cytokinesis may be responsible for the well-known rise in the incidence of binucleate and polyploid cells in the liver, with age.