I want to see the proliferation of mouse Treg cells in co-culture. Can I use soluble anti-CD3 and anti-CD28 antibodies (in place of plat bound or bead based antibodies) for Treg proliferation? and how long it must be cultured?
Thank you very much Dr. Maximiliano! Whether for mouse Treg proliferation also we need to add IL-2? If I am stimulating the cells with a pathogenic antigen then also we need to use anti-CD3 and anti-CD28 for stimulation?? Please provide if some reference is available. Thanks
Tregs can't produce IL-2, they always need a source of it for proliferation.
If you use antigen, you don't need to add a-CD3/CD28, you must include in the culture antigen presenting cells (APCs), usually irradiated whole splenocytes, purified DC, in vitro differentiated DC, etc.
Maximiliano Presa what concentration of soluble CD3 and CD28 do you use? I have been using 2ug/ml for CD3 and 1ug/ml for CD28 (without IL-2) but noticed the cell viability is quite low.