Trypsinizing MCF10A cells - (Feb/02/2007 )
ATCC recommends using soybean trypin inhibitor to inactivate trypsin after MCF10A cell release. Do I need to?
My problem: I had a confluent 60mm dish of beautiful 10A cells which I trypsinized (didn't leave on too long- about 15 min), added media (Cambrex MEGM) and spun down slowly to remove trypsin. Replated into 2 100mm dishes and they are not growing! Very few attached and after a week only a couple have divided. Did I split them at too high a ratio?
Any help would be appreciated
-nancyd-
QUOTE (nancyd @ Feb 2 2007, 09:28 AM)
ATCC recommends using soybean trypin inhibitor to inactivate trypsin after MCF10A cell release. Do I need to?
My problem: I had a confluent 60mm dish of beautiful 10A cells which I trypsinized (didn't leave on too long- about 15 min), added media (Cambrex MEGM) and spun down slowly to remove trypsin. Replated into 2 100mm dishes and they are not growing! Very few attached and after a week only a couple have divided. Did I split them at too high a ratio?
Any help would be appreciated
My problem: I had a confluent 60mm dish of beautiful 10A cells which I trypsinized (didn't leave on too long- about 15 min), added media (Cambrex MEGM) and spun down slowly to remove trypsin. Replated into 2 100mm dishes and they are not growing! Very few attached and after a week only a couple have divided. Did I split them at too high a ratio?
Any help would be appreciated
Hi!
What conc of trypsin do you use! (o,25%)
try to use 0,05%
Check you growth media (for glutamine and IGF)
Try to use insuline+transferrine+selenit mixture
Try to grown on 24 plates ( to increase cell conc )
-circlepoint-