>To the ongoing discussion about obtaining cells synchronized in the
>cycle I would like to add the following warning: The synchronization
>by transient cell arrest in the cycle induces growth imbalance and
>dramatically alters expression of cyclins and other cell cycle
>regulatory proteins. For example after double thymidine block we have
>observed "unscheduled" expression of cyclins B1 and A in cells at the
>G1/S boundary, over five-fold increase in expression of cyclin E, and
>40% increased total protein content [Gong et al., Cell Growth &
>Differentiation, 6: (November issue) 1485-93, 1995]. Kinetic and
>metabolic properties of so synchronized cells are much different
>compared to the cells from asynchronous, exponentially growing
>cultures.
>Zbigniew Darzynkiewicz
>
>
Dr. Darzynkiewicz brings up a good point. Chemical synchronization can lead
to metabolic imbalances that may invalidate or seriously compromise research
data. It is almost always preferable sychronize cells "naturally": mitotic
shake off or centrifugal elutriation.
Thomas L. Morgan, Ph.D.
Department of Radiation Oncology
Kaiser Permanente Medical Center
Los Angeles, CA 90027
morgantl@aol.com
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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