Please, Could you tell me what is the best condition (cell density/media..)to freeze and to culture human T-cells? How long can T-cells be in culture?
Thank you in advance
Mario
Mario,
Do not culture more than 1 million T-cells / mL medium. For freezing: 20 million cells / mL freezing medium (e.g. 90% FCS, 10% DMSO) is the maximum I used and 5 million the minimum. I don't know if it's the best, but it worked for me.
IJs
For freezing I think the best is 10 million/ml in 10%DMSO in FBS (better than using medium). For the culture alone 1million/ml seem reasonable and I usually take it for 48h. If they are in cocultures, for example with adherent cell they can stay more time.
What do you mean by freezing and culturing human T cells? Do you want to freeze primary T cells and then put them in culture? I wonder if it works. For Jurkat T cell line, yes you can freeze them as suggested by Paola and Ijsbrand. Human T cells, after isolation, can be cultured for 72 h max.
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