Does mast cell release its granules at physiological condition in vivo or in vitro?
If yes, what is the basal level?
i agree with Anne. In addition, medium with FCS usually induces a basal release (without upregulation of CD63) of histamine and other granule mediators
If by "physiological" you mean "no activation"... they shouldn't. In such conditions they release low levels of proteases like Tryptase-alpha and other mediators. Under IgE mediated stimulation they degranulate releasing their granule contents (preformed and stored) or, in some situations, they may release complete granules (piecemeal degranulation). This is a physiological mechanism. With laboratory handling they can degranulate by mechanic stress as stated by Anne and you need to take that into consideration and stabilize them between isolation and analysis.
I just started working in the Mast cell field and I have a question regarding the medium used to culture human primary cells since Jolien said that FCS induces basal release.
I am using RPMI 1640 + 10%FBS + L-glu + p/s + 25nM HEPES for culture and sensitization. Do you think I should use a different one to keep my cells happy? Thank you!!!!
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