We prepare mouse bone marrow-derived DC according to the protocol of Lutz (J Imm Meth 223,1999:77–92). I am in particular interested in CD86 expression and inflammatory cytokine production
I use frozen and thawed BMDC routinely for stimulation with TLR and RLR agonists (mostly nucleic acids). Usually the long-term survival is not brilliant (doesn't matter for my experiments if they survive long enough to make interferon) and endocytosis is strongly impaired (affecting e.g. endosomal TLRs and Antigen uptake).
I would only recommend it after testing that your particular readout and stimulus combination is not affected, and I always confirm results with fresh DCs afterwards (saves a little work and mice compared to always preparing fresh ones).
as another option (e.g. for precious mice), preparing DCs from frozen bone marrow works just as fine
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