Thymidine block does not synchronize L1210 mouse leukaemic cells: implications for cell cycle control, cell cycle analysis and whole-culture synchronization

It has been predicted that whole-culture methods of synchronization cannot synchronize cells. We have tested whether thymidine block, one type of whole-culture synchronization, can synchronize L1210 cells. We demonstrate experimentally that the thymidine block method cannot produce a synchronized culture. Although thymidine-treated cells are arrested primarily with an S-phase amount of DNA, there is no narrowing of the cell size distribution and there is no synchronized division pattern following release from the thymidine block. In contrast to a whole-culture synchronization method, cells produced by a selective (i.e. non-whole-culture) method not only have a specific DNA content, but also have a narrow size distribution and divide synchronously. Generalizing the results to other cell lines, we suggest that these conclusions call into question experimental measurements of gene expression during the division cycle based on thymidine inhibition synchronization.