Chapter 13 Cell Synchronization

Publisher Summary

This chapter describes the methods of analyzing cell-cycle kinetics, methods for synchronizing mammalian cells, and caveats in interpreting cell synchrony experiments. Many cell-synchronization protocols involve placing cells under growth inhibitory conditions for periods of time related to the duration of specific cell-cycle phases. The synchronization methods include release from GO arrest, release from M- and S-phase blocking agents, mitotic detachment, and centrifugal elutriation. To distinguish between growth-dependent and cell cycle-dependent regulation, cells are generally synchronized using more than one method. In some cases, the use of two synchronization methods reveals that all or part of an observed biochemical change is not cell-cycle dependent. The chapter also describes methods for analyzing the cell-cycle kinetics of asynchronous mammalian cells. The asynchronous cell methods include the determination of mean generation time and proliferative fraction, the determination of mitotic and [³H]thymidine-labeling indices, and the estimation of cell-cycle phase durations by the labeled mitoses procedure and by flow cytometry.