I am new to immunology field and have a question as a novice: If I want resting NK cells as controls in vitro, can I grow the NK cells without cytokine stimulation in vitro? only for a day (overnight incubation). My only aim in doing this is to use it as a control without stimulation and compare it with NK cells that I will be activating using IL-2 and IL-15.
Overnight incubation is OK.
They need cytokine for long term incubation/proliferation in culture.
For overnight incubation, I will suggest to put at least 1ug/ml of IL-15 to maintain them alive and avoid stimulation of apoptotic mechanism. Are you studying isolate NK cells or bulk PBMC ?
Maryam Mehrabi Thank you !
Olivier Lucar Thank you for your suggestion. I am interested in isolating only the NK cells from PBMC by negative selection.
Ok so yes you should add at least IL-15 because isolated NK cells would start to die without a survival factor
In my experience, overnight incubation of primary NK cells, freshly isolated by negative selection from peripheral blood is unproblematic. It is different matter of course, if they have been exposed to harsher isolation procedures (e.g. isolation from tissues or cell sorting). Cytokines should be dispensable for such a short time period and the intended use but you could opt for more fancy medium to be on the safe side. Good luck!
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