After freezing stock and thawing as per the recommendations, NK cells have many dead cells. Not able to get rid of dead cells, any suggestions
HI,
· Freezing and thawing procedures are stressful to most cells. Do notvortex, bang the flasks to dislodge the cells (except when culturing insect cells), or centrifuge the cells at high speeds.
· Make sure that you thaw the frozen cells quickly, but dilute them slowly using pre-warmed growth medium before plating.
· Allow the culture more time to recover. Sometimes it may take a few weeks for the cells to form a monolayer or dense suspension, depending on the age and passage number of the cells when frozen or position in the growth phase. Optimum conditions to freeze are when cells are in the log phase.
all the best
Some additional information about the line would be useful. Some NK cell lines need special culture conditions, suchs as adding IL2 to the culture media, as in the case of the NKL or NK92 cell lines. You should check that. Also, be careful when removing the culture media, you could be taking the most viable cells from the top, since the reduced granularity.
The NK cells are NK92 and they are suspension cells, grow mostly in clumps. I am adding IL-2 as recommended from the company.
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