EDTA to weaken cell attachement - (Feb/07/2007 )
Hi!
I am planning to weaken the bonds of a confluent layer of cells by adding some EDTA for about a minute (eventually I require a cell sheet). As the whole system is serum-free, I cannot use media with serum to stop the action of the EDTA (which would prevent most of the cells from lifting off). Would PBS do the job? After the treatment I will be placing the cells in serum free media again.
Thanks
-Shefmat-
QUOTE (Shefmat @ Feb 7 2007, 12:07 PM)
Hi!
I am planning to weaken the bonds of a confluent layer of cells by adding some EDTA for about a minute (eventually I require a cell sheet). As the whole system is serum-free, I cannot use media with serum to stop the action of the EDTA (which would prevent most of the cells from lifting off). Would PBS do the job? After the treatment I will be placing the cells in serum free media again.
Thanks
I am planning to weaken the bonds of a confluent layer of cells by adding some EDTA for about a minute (eventually I require a cell sheet). As the whole system is serum-free, I cannot use media with serum to stop the action of the EDTA (which would prevent most of the cells from lifting off). Would PBS do the job? After the treatment I will be placing the cells in serum free media again.
Thanks
PBS or your serum-free medium will do; it will be a stop of EDTA-chelating action by dilution, washing out, and, in the case of medium, offering of bivalent cations; serum-containing medium is more useful to stop trypsin (of a trypsin-EDTA solution)
-The Bearer-