2 replies to this topic

[OKSO]

Hi all,

I wonder if anyone can tell me what the best conditions to differentiate U937s are?

Looking at a few papers the concentration ranges from 10nM to 160nM PMA...
Incubation periods range from 12h to 48 hours - has anyone stimulated for 72h?

Also what is a good starting point for cell density - some people use 1xE5/ml others 7xE5/ml

Thanks so much in advance!!

[SuMi]

I use 50 ng/ml PMA. I seed the U-937 cells at 1e6 cells/ml (3e6 per well of 6-well plate) and add the PMA for 24 hours. I then remove the medium and add fresh medium without PMA and let the cells culture for a further 48 hours (i.e. 72 hours total). Some people don't remove the PMA but I prefer to not have my cells over-stimulated.

[OKSO]

Thanks for the information SuMi! Cheers

SuMi, on Mar 4 2010, 02:09 PM, said:

I use 50 ng/ml PMA. I seed the U-937 cells at 1e6 cells/ml (3e6 per well of 6-well plate) and add the PMA for 24 hours. I then remove the medium and add fresh medium without PMA and let the cells culture for a further 48 hours (i.e. 72 hours total). Some people don't remove the PMA but I prefer to not have my cells over-stimulated.