Hello
It is been a long time I have been trying to stimulate vascular smooth muscle cells (MOVAS) for ROS generation. I am using DCFDA assay for ROS detection....
Everything I tried, has failed to stimulate it! LPS (short and long incubation time), high and low concentration!, Iron (Fe3+), glucose (25 mM 50 mM etc), Beta-glycerophosphate (up to 10 mM).... none works!
Does anybody have any idea what is happening?
Extremely frustrating cycle of trial and failure !!!
If you need a positive control, just treat them with H2O2. Otherwise nicotine or high calcium (3.6 or 5.4 mM) worked in my hands, in primary human vascular smooth muscle cells.
The problem might be in the MOVAS cells themselves? Maybe the fact that it's a cell line makes them insensitive to stimuli.
Gosia Furmanik Thanks very much for your prompt reply.
I will give calcium supplementation a try.
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