After PBMC isolation, we are not able to activate and expand T cells.
Dear Dr. Payal Agarwal
After isolation of PBMCs, you may activate the T cells in culture by adding mitogens (which are potent stimulators of T-cell activation and proliferation). For instance, PHA or concanavalin A which are mitogenic lectins are polyclonal activators, in that they activate lymphocytes, irrespective of their antigenic specificity.
Or
You may use monoclonal antibodies, for example, anti-CD3/CD28, which provides a co-stimulatory signal that engages the TCR, which can be used for antigen-induced activation.
Also, specific cytokines, such as interleukin-2 is added to culture to support T cell expansion and survival.
You may use 1-5 µg/ml of PHA along with 100-300 IU/mL of IL-2 to efficiently activate T cells and enhance T-cell cluster formation.
Regards,
Malcolm Nobre
T-cell activation can be achieved by 5 ug/ml Concanavalin A in a timely exponential increase up to 60 min. of incubation of PBMC suspensions.
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