Cell death after PMA treatment - (Aug/07/2008 )
Hi,
I am trying to activate my Jurkat cells with PMA with different concentration of 25, 50, 100 ng. But they stary dying. I am not sure if it is usual or not.
I am trying to activate my Jurkat cells with PMA with different concentration of 25, 50, 100 ng. But they stary dying. I am not sure if it is usual or not.
absolute amounts do not say much, and the incubation times are missing; what is meant by "to activate my Jurkat cells? what are "activated " Jurkat cells?
my friends use PMA to induce U937 into monolayer.
she tried out different concentration and monitor how long it takes to make maximun number of cells induced and stay viable.
i think you might need to optimise on that
We tend to use PMA at about 1ng/mL for peripheral blood cells in a 10mL volume.
PHA could be an alternative to use at 1ug/mL.
How long does it take for them to start dying? What media are they in?
Hi,
Stimulation with 10 ng/ml of PMA and 1 µM of IO for 4 hours woked for me. Furthermore for 24 Hrs and 48 Hrs culturing I washed the cells after 4 hours of stimulation and continue the culture with fresh medum without PMA/IO. In the literature, you could have noticed that various authors had used 50 ng/ml of PMA and 5 µM of IO ,just to make sure that all the cells gets activated so that these cells can be used for FACS studies. In the mean time, it would be of interest to know about the clone of Jurkat cells you used and what are the cytokines you measured?
Good luck in your research
hi, I am using 20ng/ml PMA to acitvate my monocytic cell lines for 24 hours. It works very well. I think you'd better use PHA+IL-2 to activate the T cell line.