Hello,
I'm trying to infect bone marrow dendritic cells with M. bovis at different M.O.I. Whats the recommended and maximum amount of time I should allow the infection before it kills the BMDC? Plus, can I use the spinfection (infect by spinning)? If so, what are the parameters. Thank you.
M.bovis, is the virulent strain right, not BCG? Either way, it depends on what you want to measure.
Even virulent strains, you can do a MOI at 1:10 (1 bacterium:10 cells) and the cells are usually viable for quite a while. If you do a 10:1, then you will start see cell death around 72hrs.
If you want to do any intracellular measuring of bacteria growth, activity, etc, then the rule is infect for 4 hours and wash away extracellular bacteria. And then culture for whatever time point you want.
What do you mean infection by spinning? The BMDCs should already be adherent, and the bacteria will also settle, upon which the BMDCs should then phagocytose the bacteria.
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