We would like to keep in culture freshly-isolated mouse memory CD4 T cells. We are currently trying to keep them with complete RPMI + 10% serum supplemented with IL-2 (100 U/ml), but within 24h there is a significant drop in the population.
Hi Giorgio. If you are dealing with real memory T cells just leave them in culture and do not give them IL-2. Memory T cells become quiescent and do not divide unless there is their antigen and signal II is present. Otherwise they can stay in culture for a long time. Have a look at our papers attached.
Hi Giorgio. If you are dealing with real memory T cells just leave them in culture and do not give them IL-2. Memory T cells become quiescent and do not divide unless there is their antigen and signal II is present. Otherwise they can stay in culture for a long time. Have a look at our papers attached.
Scheherazade and Celestina, thank you so much for your prompt responses. Our issue is that we do not want to add any stimulation to the cells (like PHA) as we want to measure their response to specific co-stimulatory signals. Scheherazade, thank you so much for the articles. Just one clarification: I do not see what conditions (type of media and growth factors) you cultured the Tmem in. In our hands, if we do not add anything (just RPMI/10% serum), approximately 80% of the cells are dead in 2-3 days based on viability staining...
That is exactly what you want. Only memory cells remain and the other cells die off. The papers I attached show that memory cells do not proliferate and are quiescent. The culturing conditions is not in those papers. It is unpublished data.
Hi Giorgio,
Have you tried adding IL-7 instead of IL-2? I know naive T cells survive a lot better in the presence of IL-7, and given that memory cells should express the IL-7 receptor it's worth a shot.
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