Hi For the CD8+ CD28- T cell expansion ( terminally differentiated memory T cells), in addition to the anti-cd3, what cytokines or other substances should i add?How about IL-2, IL-15, T+I or IL-7 ?
Thanks in advance for your advice
You might be already doing it but your anti-CD3 must be plate-bound so that you can cross-link the TCR. I would suggest to use PHA and all the cytokines you mentioned: IL-2, IL-7, IL-15. After 2-3 days remove the media and replace with PHA free media (but containing all cytokines).
No worries. I should've mentioned that this protocol is quite robust for expanding human T cells, it might be sligthly different for mouse T cells. You could also consider using irradiated APCs as stimulators. Hope it works for you!
I have another question. First cultured PBMC and after that sorted them or first sorted and after that cultured them? Can you sure me whether culturing PBMC and after that sorting them is better or sorting before the culturing?
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