Hi again!
I've just started growing CTLL-2 IL2 dependent T cells in culture, and need quite high numbers.
The ATCC recommendations for culture are to subculture actively growing cells BEFORE they reach 2x10-5 cells/ml, but for my purposes this doesn't give me as many as I would like.
I just wondered if anyone has experience maintaining these cells above this density, perhaps to 5x10-5/ml?
I have plated them epxerimentally at 1x10-6/ml for 24 hours with viable, happy cells.
Thanks for your input :)
Brittany
Hi Brittany,
I don't have any experience using these particular cells, but in general you can easily culture T cells in IL-2 containing medium up to ~1.5x10e6 cells/mL. Just make sure you use a medium with a strong buffer capacity to maintain acceptable pH, such as RPMI with 25 mM HEPES.
Good luck,
Jeroen
Hi Jeroen,
Thanks for getting back to me. It will be a case of trial and error I think! Thanks for the advice - that's encouraging :)
Best
Brittany
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