I'm planning on activating 5 million PBMCs with CD3/CD28 at the same time applying a treatment, then incubate for 72 hours. Then I seperate cell subtypes using antibody sequencial seperation method. I initially thought of plating them in 150mm non-treated plates but now I'm second-guessing because many literature indicates plating PBMCs at 1 million cells per mL in 24 well plates or 6-well plates. will 150mm plates be too big and can affect the viability of the cells?
Usually we plate 1.5 million of cells per well in 6 well plates (1 million cells/ml), if you want to plate 5 million of cells, I will use T25 culture flasks, which might be more convenient
HI! l agree with Dominique Duval
For this number of PBMC, I usually worked in T25 flasks with 5ml of medium
