Synchronization of mouse L-P59 cells by centrifugal elutriation separation

Cell separation by centrifugal elutriation was employed to obtain synchronized cells from asynchronous populations of mouse L-P59 fibroblast cells. The fractions most enriched in specific phases of the cell cycle contained about 90% G1-phase cells, 70% S phase cells and 60% G2 + M phase cells, respectively, as determined by pulse cytophotometry and autoradiography. When replated, cells from separated fractions divided synchronously in culture. The method is rapid and large numbers of cells (3 × 10⁸) can be processed in a single run with no loss of viability. This method should be applicable to most cell lines which can be obtained in single-cell suspension.