Protocol tip:Optimal concentration range of antibodies anti-CD3 and anti-CD28 should be determined empirically. For anti-CD3 (mouse and human), optimal concentration range is 1–3 µg/mL and for anti-CD28 (mouse and human), optimal concentration range is 3–5 µg/mL.
IL-2 stimulates the growth and differentiation of cytotoxic T cells. Adding 100 ng
IL-2 recombinant protein
(Cat. No. PHC0026) instead of anti-CD28 can help expand this population.
Protocol tip:Anti-CD3 can be directly added to the media in step 3 at a final concentration of 1 µg/mL to save time.
This protocol can be scaled for larger cell quantities by using