synchronization of cell lines
gollin at MED.PITT.EDU
Fri Apr 14 07:44:14 EST 1995
It is notoriously difficult to synchronize cell lines. One reason may be that
p53 abnormalities abrogate cell cycle checkpoints, as elegantly discussed in
Cross et al. Science 267:1353-1356 (1995). Other checkpoints are likely to
be altered as well in cell lines, since cells that grow well in culture may
have been selected for on the basis of cell cycle abnormalities.
One may try the usual methotrexate synchronization protocol used for high
resolution chromosome studies (speak to your local cytogeneticist for the
protocol), but this may not work in these cell lines. The reason that
colcemid may not work is clearly shown in the Cross et al. paper. This
is a very frustrating problem for cytogeneticists when we are observing
cell division and not obtaining many metaphases in our preparations. So
I sympathize. But that doesn't help. Gerald Holmquist and I spent many days
trying to synchronize a lymphoblastoid line in the early 1980's and the
best success was nitrous oxide under pressure, a technique used by Potu
Rao and Walter Hittelman in the 80's to make prematurely condensed chromosomes.
Best of luck.
Susanne Gollin, Ph.D.
Assistant Professor of Human Genetics
University of Pittsburgh and the Pittsburgh Cancer Institute
gollin at med.pitt.edu
More information about the Cellbiol