Dear all,
I started performing Mixed Lymphocyte Reaction (MLR) to observe the immunomodulatory effects of regulatory T cells. The protocol stimulates the lymphocytes from the mouse spleen using anti-CD3 and anti-CD28 beads but didn't work very well. My positive control proliferated in the same way as my negative control. Unexpectedly., my condition treated with Tregs proliferated more than others conditions and there were more viable cells as well. Density 1x10E5/well P96 flat bottom. I tested different culture times, 2,3,4,6 days all with the same results. But when I tested with the density of 3x10E5 the results were better. I already tested one condition with PHA to stimulate.
So I was thinking to co-culture mouse splenocytes as donor 1 and human PBMC as donor 2 to perform MLR.
Has anyone done that or have some tips?
All the best,
Did you ever do the MLR - Co-culture Mouse splenocytes and PBMC human?
I also want to this for prediction of GVHD
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