Forum for prokaryotic and eukaryotic culture, cell culture related assays (e.g. CTL, cytokine release, B cell culture), cell lines/primary culture, reagents, etc.
I'm trying to get an NK-cell assay working and have so far tried an LDH release assay and also by FACS using CFSE/PI. Problem is that I have yet to see any target cell lysis. Is it possible that YAC-1 cells lose their sensitivity to lysis after prolongued periods in culture? I thaw my cells and use them within 3 passages so they don't exactly spend a long time in culture...
I've also tried stimulating the isolated spleen cells with interferon and saw no appreciable difference in cell death. I would go so far as to say I saw not a single dead target cell at an E:T ratio of 100:1, which according to the literature should give me ~10% death in unstimulated cells and >40% in IFN induced NK cells.
Does anyone have any comments? Should I look at my target cells?
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