I separated the spleen mononuclear cells (SMCs) from the C57 mouse spleen by using a Ficoll and seed 800,000 cells per well at volume 500 microliter and incubated for48 hours. Is the amount of the cytokines so large that I can measure cytokine by Eliza method? or I must use mitogens for stimulation of spleen mononuclear cells or change incubation time or cells.
(I prescribed medication to some groups, and I expect cytokine production to be different in treated and untreated groups)
What Cytokine you are interested in. Under steady state these cells won't produce much of the cytokines. You might have to challenge these cells for cytokine production.
Another thing how would you expect that this medication is going to affect the cytokine production when you culture them in vitro.
I want to measure IL-1 and 6 and 27 in the culture supernatant. I inject anti-inflammatory drugs to mice and my expected is that this cytokine decrease in drug injected mouse groups.
It is better to look for the serum cytokines and challenge mice rather than looking cytokines in vitro. Hope this will help you
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