Hello, world!
Have anyone ever expanded human or murine CD4+ T cells without CD3/CD28 beads? Is it possible?
We are triing to do it using RPMI-1640 with glutamine, ME, Il-2 on the plates covered with aCD3 antibodies and with addition of aCD28 antibodies in the medium. The cells clearly are activated, but their numbers grow slowly or even decrease. No one in our lab have ever done such thing, and I can't find good protocol. Or maybe I'm wrong and beads are mandatory?
Hi
The anti-CD3/CD28 antibodies can also be used to activate T cells.
Check the amount of antibody and IL-2 you are using to activate T cells.
Use sub-optimal stimulation which will prevent AICD of your cells.
Hi,
I'm just starting in the lab and using this technique to activate CD4+ human T cells too. My protocol has 25U/ml IL-2. I am now ordering my own reagents. Apparently the IL-2 is easily found & commercially available everywhere, anyone know which one I should order in UK?
Hello,
To activate T cells, it's possible to use planar artificial "target surface", an optical glass covered with antiCD3 antibody; other suitable combinations of antibodies can be employed, too.
Bunnell, S. C., V. A. Barr, C. L. Fuller, and L. E. Samelson. 2003.
High-resolution multicolor imaging of dynamic signaling complexes in T cells stimulated by planar substrates. Sci. STKE. 2003:pl8.
https://www.researchgate.net/requests/r25322050
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