synchronization of cell lines

[Dirk Dittmer]

In article <h-petrie-1304951121570001 at mac-133.k-g2.ski.mskcc.org>,
h-petrie at ski.mskcc.org (Howard T. Petrie) wrote:

> We have been trying for some time now to synchronize (preferably in M
> phase) several murine cell lines that we work with, two lymphoid and one
> myeloid.  We have extensively tried protocols using colcemid and
> Nycodazole, as well as limited attempts with other approaches.  The
> problem we are experiencing is that any concentration which is apparently
> effective at blocking mitosis is also toxic.  We have tried titrations of
> both time and drug concentration without improvement.  The benefit of
> anyone else's experience would be greatly appreciated


I have tried mitotic shake off with and without nocodazole and it worked
nicely with NIH3T3 cells. You can isolate approx. 1/2 of the starting
population that way. However, you need cells that grow attached.
The other way would elutriation. 

Dirk