All protocols are based on cell concentrations which makes sense for nutrients and else supplies but one must probably take into account the density of the cells after they have sedimented at the bottom of the up-stood flask so they don't "suffocate", I assume.
Though, I cannot find anywhere some information about the recommended maximum number of PBMC per cm2. Would anyone have some expertise about this? Thanks!!!
I would recommend you not to exceed 5 * 10E6 cells per cm2. About it logarithmic growth is often not safe. In the upright position, it is useful to shake the bottles gently every 12 hours.
I want to have cell to cell virus transmission so I'm not sure I will disturb the cells as much as possible. They should not proliferate so I will go for 5x10e6 max per cm2. Thanks a lot :)
PS where do you have this number from? Wild guess, experience or literature?
I have worked with HI virus. In my PhD, a detailed procedure is described. Good Luck!
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