I'm planning to incubate mouse T cells with my agent of interest for 7 days. I want to avoid cells reached confluency at a point that affects viability. However, I want to have enough cells to do flow at the end of the incubation period. Which plates work best and how many CFSE-stained cells should I seed? I'm planning to coat the plate with anti-CD3 (1ug/mL) and add soluble anti-CD28 (5ug/mL) to the medium for T cells activation. Also, I'll change the media at day 4 of incubation. Any practical tip when changing the media, to lose as few cells as possible?

I'll appreciate any comment. Thanks a lot!