Hello everyone,
lately we experienced some problems with the performance of our NK92 cells, because they are performing way worse in cytotoxic assays than a few months ago. We thought this might either result from the NK92 cells being exhausted or resting. So we tested for the NK cell exhaustion markers DNAM-1 and NKG2a. Our NK92 cells were negative for DNAM-1 and positive for NKG2a, which might indicate that they`re in an inactive state.
Has anyone else experienced this before? Or does anyone know how to reverse exhaustion in NK92 cells?
*Medium + 100U/ml is given two-three times a week
Hi,
your description suggests that you culture your line for a few months. In my experience, most of the popular cell lines are in general "exhausted" after ~3 months of culture, so normally I keep the cells for 2-2.5 months in culture maximum, then I thaw a new batch. I haven't experienced such exhaustion in NK-92 cells, but that was also the line which I cultured for ~2 months.
Another thing is inositol, I was also adding it fresh in the culture, final concentration was 0.2 mM in the media. I kept IL-2 and inositol frozen in -20C for longer periods of time, and when I thawed an aliquot, I kept it in 4C for a maximum 1 week.
Of course, those are my suggestions regarding cell culture, have a look and compare with your routine. I can provide full media formulation for NK-92 that worked for me on demans.
Good luck!
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts