Centrifugal elutriation as a means of cell cycle phase separation and synchronisation.
The ability to purify cells in specific phases of the cell cycle in sufficient quantities for biochemical analysis or to obtain a "synchronized" population of living cells enriched in a particular phase is of great value for studying a wide range of cell cycle processes. Transformed tumour cell lines, such as DT40, can be arrested and released from specific points in the cell cycle using drugs which inhibit DNA or protein synthesis or interfere with mitotic spindle function, however such drugs are frequently toxic and may themselves perturb the cell cycle process or phenomenon under study. Centrifugal elutriation provides a means of separating an unperturbed culture of living cells into highly enriched G1-, S-, and G2/ M-phase fractions. The resulting cell fractions can be analysed directly for protein and mRNA expression or returned to culture and their subsequent progression through the cell cycle monitored by flow cytometry, microscopy, or other techniques.
Gillespie
David A F
DA
CR-UK Beatson Laboratories, Beatson Institute for Cancer Research, Garscube Estate, Bearsden, Glasgow G61 1BD.
Henriques
Catarina
C
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D002453:Cell Cycle; D002460:Cell Line; D002498:Centrifugation; D002645:Chickens