synchronization of primary human cells

From: Wojciech Niedzwiedz (wniedzwiedz@yahoo.com)
Date: Thu May 24 2001 - 03:23:54 EST


Hi,
I was wondering if anyone could help me. I am trying
to synchronize amniotic fluid cells in G1, S and G2
phases of the cell cycle. I have never done this
before and I do not have a working protocol to start
with. Till now I've tried the serum withdrawal for 48
h for G0/G1, and for S phase - 48h of serum starvation
followed by 15 h of incubation with aphidicolin (6uM);
serum withdrawal allowed me to get 85% of cells in
G0/G1, the method for S phase synchronization I used
yielded in 75% of cells at S phase. I would really
appreciate if someone could provide me with a protocol
that would give a better efficiency of synchronization
with regards to amniotic fluid cells.

Thank you in advance,
Wojciech Niedzwiedz
University of Bristol
AOC Orthopaedic Center


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