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T84s and interferon gamma sensitivity

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#1 kanter



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Posted 16 February 2011 - 05:46 PM

Hi all,
me and my T84s again.:(
The cells are growing well on PET membranes, however I've run into a different problem: the cells seem to have lost sensitivity to IFNgamma.
Does anyone know why this may happen? Would passage number, possible mycoplasma infection, or anything else play a role?
I have used a couple of different cell batches, with different passage numbers, and both don't respond- whereas when I started the experiment the cells responded well, dropped TER about 4 fold in 48 hours.
My lab doesn't test for mycoplasma, which is why I'm asking about this too.
The IFNgamma has a short storage life when reconstituted, 3 months, and it's been about 2.5- could it have gone off early??
Is seeding density crucial- for example is cells overgrow (eg. "climb" on top of each other) will they become insensitive to IFN??
Any ideas???
Thanks so much!

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