Mechanistic insight into the suppressive effects of stem cell factor during IgE-mediated activation in bone marrow-derived mast cells.

Abstract

Abstract Treatment of bone marrow derived mast cells (BMMCs) with stem cell factor (SCF) is common practice when differentiating mast cells for experimentation. The enhanced growing potential from including SCF during mast cell culture and differentiation allows for easier maintenance and quicker experiments. Recent publications have begun to identify potentially negative effects of treatment with SCF regarding mast cell activation. The mechanism of SCF inhibition on mast cell activation is mostly unknown, but BMMCs derived with the addition of SCF function differently than untreated cells when mast cells are primed and subsequently activated. This study provides insight into the specific targets of SCF treatment and the impact SCF has on the ability of mast cells to activate. Running ELISAs in conjunction with real-time qPCR, we identified differentially expressed proteins involved in FCɛRI signaling and mast cell activation in BMMCs exposed to SCF. We examine the effects of SCF on Fyn, Lyn, Syk and Stat5 protein expression compared to mast cells raised without SCF. These data are also placed in the context of other immunosuppressive mechanism, (e.g. TGFβ). The presented data allow researchers to better understand the implications that may result from SCF treatment of BMMCs, especially with respect to examining IgE-mediated functions.